Diazole amides

ABSTRACT

Compounds are provided that act as potent antagonists of the CCR1 receptor, and have in vivo anti-inflammatory activity. The compounds are diazole lactam derivatives and are useful in pharmaceutical compositions, methods for the treatment of CCR1-mediated disease, and as controls in assays for the identification of competitive CCR1 antagonists.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/878,784 filed Oct. 8, 2015, which application is a divisional of U.S.patent application Ser. No. 14/137,479 filed Dec. 20, 2013 (now U.S.Pat. No. 9,169,248), which application claims the benefit of priority toU.S. Provisional application Ser. No. 61/745,444, filed Dec. 21, 2012,each of which is incorporated herein by reference.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

Not Applicable

BACKGROUND OF THE INVENTION

The present invention provides compounds, pharmaceutical compositionscontaining one or more of those compounds or their pharmaceuticallyacceptable salts, which are effective in inhibiting the binding ofvarious chemokines, such as MIP-1α, leukotactin, MPIF-1 and RANTES, tothe CCR1 receptor. As antagonists or modulators for the CCR1 receptor,the compounds and compositions have utility in treating inflammatory andimmune disorder conditions and diseases.

Human health depends on the body's ability to detect and destroy foreignpathogens that might otherwise take valuable resources from theindividual and/or induce illness. The immune system, which comprisesleukocytes (white blood cells (WBCs): T and B lymphocytes, monocytes,macrophages granulocytes, NK cell, mast cells, dendritic cell, andimmune derived cells (for example, osteoclasts)), lymphoid tissues andlymphoid vessels, is the body's defense system. To combat infection,white blood cells circulate throughout the body to detect pathogens.Once a pathogen is detected, innate immune cells and cytotoxic T cellsin particular are recruited to the infection site to destroy thepathogen. Chemokines act as molecular beacons for the recruitment andactivation of immune cells, such as lymphocytes, monocytes andgranulocytes, identifying sites where pathogens exist.

Despite the immune system's regulation of pathogens, certaininappropriate chemokine signaling can develop and has been attributed totriggering or sustaining inflammatory disorders, such as rheumatoidarthritis, multiple sclerosis and others. For example, in rheumatoidarthritis, unregulated chemokine accumulation in bone joints attractsand activates infiltrating macrophages and T-cells. The activities ofthese cells induce synovial cell proliferation that leads, at least inpart, to inflammation and eventual bone and cartilage loss (see,DeVries, M. E., et al., Semin Immunol 11(2):95-104 (1999)). A hallmarkof some demyelinating diseases such as multiple sclerosis is thechemokine-mediated monocyte/macrophage and T cell recruitment to thecentral nervous system (see, Kennedy, et al., J. Clin. Immunol.19(5):273-279 (1999)). Chemokine recruitment of destructive WBCs totransplants has been implicated in their subsequent rejection. See,DeVries, M. E., et al., ibid. Because chemokines play pivotal roles ininflammation and lymphocyte development, the ability to specificallymanipulate their activity has enormous impact on ameliorating andhalting diseases that currently have no satisfactory treatment. Inaddition, transplant rejection may be minimized without the generalizedand complicating effects of costly immunosuppressive pharmaceuticals.

Chemokines, a group of greater than 40 small peptides (7-10 kD), ligatereceptors expressed primarily on WBCs or immune derived cells, andsignal through G-protein-coupled signaling cascades to mediate theirchemoattractant and chemostimulant functions. Receptors may bind morethan one ligand; for example, the receptor CCR1 ligates RANTES(regulated on activation normal T cell expressed), MIP-1α (macrophageinflammatory protein), MPIF-1/CKβ8, and Leukotactin chemokines (amongothers with lesser affinities). To date, 24 chemokine receptors areknown. The sheer number of chemokines, multiple ligand bindingreceptors, and different receptor profiles on immune cells allow fortightly controlled and specific immune responses. See, Rossi, et al.,Ann. Rev. Immunol. 18(1):217-242 (2000). Chemokine activity can becontrolled through the modulation of their corresponding receptors,treating related inflammatory and immunological diseases and enablingorgan and tissue transplants.

The receptor CCR1 and its chemokine ligands, including, for exampleMIP-1α, MPIF-1/CKβ8, leukotactin and RANTES, represent significanttherapeutic targets (see Saeki, et al., Current Pharmaceutical Design9:1201-1208 (2003)) since they have been implicated in rheumatoidarthritis, transplant rejection (see, DeVries, M. E., et al., ibid.),and multiple sclerosis (see, Fischer, et al., J Neuroimmunol.110(1-2):195-208 (2000); Izikson, et al., J. Exp. Med. 192(7): 1075-1080(2000); and Rottman, et al., Eur. J. Immunol. 30(8):2372-2377 (2000). Infact, function-blocking antibodies, modified chemokine receptor ligandsand small organic compounds have been discovered, some of which havebeen successfully demonstrated to prevent or treat somechemokine-mediated diseases (reviewed in Rossi, et al., ibid.). Notably,in an experimental model of rheumatoid arthritis, disease development isdiminished when a signaling-blocking, modified-RANTES ligand isadministered (see Plater-Zyberk, et al., Immunol Lett. 57(1-3): 117-120(1997)). While function-blocking antibody and small peptide therapiesare promising, they suffer from the perils of degradation, extremelyshort half-lives once administered, and prohibitive expense to developand manufacture, characteristic of most proteins. Small organiccompounds are preferable since they often have longer half lives invivo, require fewer doses to be effective, can often be administeredorally, and are consequently less expensive. Some organic antagonists ofCCR1 have been previously described (see, Hesselgesser, et al., J. Biol.Chem. 273(25): 15687-15692 (1998); Ng, et al., J. Med. Chem.42(22):4680-4694 (1999); Liang, et al., J. Biol. Chem.275(25):19000-19008 (2000); and Liang, et al., Eur. J. Pharmacol.389(1):41-49 (2000)). In view of the effectiveness demonstrated fortreatment of disease in animal models (see, Liang, et al., J. Biol.Chem. 275(25): 19000-19008 (2000)), the search has continued to identifyadditional compounds that can be used in the treatment of diseasesmediated by CCR1 signaling.

BRIEF SUMMARY OF THE INVENTION

The present invention relates to compounds having formula I:

or a pharmaceutically acceptable salt, hydrate, solvate, N-oxide orrotamer thereof. In Formula I, each A is independently selected from thegroup consisting of N and CH;

-   X and Z are each independently selected from the group consisting    -   (i) monocyclic or fused-bicyclic aryl and heteroaryl, wherein        the heteroaryl group has from 1-4 heteroatoms as ring members        selected from N, O and S;    -   (ii) monocyclic four-, five-, six- or seven-membered ring        selected from the group consisting of cycloalkane, and        heterocycloalkane, wherein the heterocycloalkane rings have from        1-3 heteroatoms as ring members selected from N, O and S;    -   wherein each of the rings in (i) and (ii) are optionally        substituted with from 1 to 5 substituents selected from halogen,        CN, C₁₋₈ alkyl, C₃₋₈ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,        C₁₋₈ haloalkyl, C₁₋₈ hydroxyalkyl, —OR^(a), —CO₂R^(a),        —SO₂R^(a), —NR^(a)R^(b), —CONR^(a)R^(b), aryl, 5- or 6-membered        heteroaryl, and 3-, 4-, 5- or 6-membered heterocycloalkane        wherein the heteroatoms present as ring vertices of the        heteroaryl and heterocycloalkane rings are selected from N, O        and S, and wherein the alkyl, cycloalkyl, aryl, heteroaryl and        hetereocycloalkane portions of the substituents are optionally        further substituted with 1-3 R^(a); and optionally, two        substituents on adjacent ring vertices are connected to form an        additional 5- or 6-membered ring which is saturated, unsaturated        or aromatic having ring vertices selected from C, O, N and S;    -   R³ is a member selected from the group consisting of H, halogen,        CN, C₁₋₈ alkyl, C₃₋₈ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,        C₁₋₈ haloalkyl, C₁₋₈ hydroxyalkyl, —OR^(a), —CO₂R^(a),        —NR^(a)R^(b), —CONR^(a)R^(b), aryl, 5- or 6-membered heteroaryl,        and 3-, 4-, 5- or 6-membered heterocyclic wherein the        heteroatoms present as ring vertices of the heteroaryl and        heterocyclic rings are selected from N, O and S, and wherein the        alkyl, cycloalkyl, aryl, heteroaryl and hetereocyclic portions        of R³ are optionally further substituted with 1-3 R^(a);    -   R⁴ is a member selected from the group consisting of H, —OR^(a)        and C₁₋₈ alkyl optionally substituted with —OR^(a) or        —NR^(a)R^(b); or R⁴ is combined with X to form a bicyclic fused        ring system;        each R^(a) and R^(b) is independently selected from the group        consisting of hydrogen, hydroxyl, halogen, cyano, C₁₋₈ alkyl,        C₁₋₈ alkoxy, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₃₋₆        cycloalkylalkyl, amino, C₁₋₈ alkylamino, di C₁₋₈ alkylamino,        carboxamide, carboxy C₁₋₄ alkyl ester, carboxylic acid, and        —SO₂—C₁₋₈ alkyl.

In addition to the compounds provided herein, the present inventionfurther provides pharmaceutical compositions containing one or more ofthese compounds, as well as methods for the use of these compoundsprimarily to treat diseases associated with CCR1 signalling activity.

BRIEF DESCRIPTION OF THE DRAWINGS

None

DETAILED DESCRIPTION OF THE INVENTION

I. Abbreviation and Definitions

The term “alkyl”, by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, having the number of carbon atoms designated (i.e. C₁₋₈ meansone to eight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. The term “alkenyl” refers toan unsaturated alkyl group having one or more double bonds. Similarly,the term “alkynyl” refers to an unsaturated alkyl group having one ormore triple bonds. Examples of such unsaturated alkyl groups includevinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “cycloalkyl”refers to hydrocarbon rings having the indicated number of ring atoms(e.g., C₃₋₆cycloalkyl) and being fully saturated or having no more thanone double bond between ring vertices. “Cycloalkyl” is also meant torefer to bicyclic and polycyclic hydrocarbon rings such as, for example,bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term“heterocycloalkane” or “heterocycloalkyl” refers to a cycloalkyl groupthat contains from one to five heteroatoms selected from N, O, and S,wherein the nitrogen and sulfur atoms are optionally oxidized, and thenitrogen atom(s) are optionally quaternized. The heterocycloalkane maybe a monocyclic, a bicyclic or a polycylic ring system. Non limitingexamples of heterocycloalkane groups include pyrrolidine, imidazolidine,pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin,dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine,thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide,piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone,tetrahydrofuran, tetrhydrothiophene, quinuclidine, and the like. Aheterocycloalkane group can be attached to the remainder of the moleculethrough a ring carbon or a heteroatom.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified by—CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group will have from 1to 24 carbon atoms, with those groups having 10 or fewer carbon atomsbeing preferred in the present invention. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingfour or fewer carbon atoms. Similarly, “alkenylene” and “alkynylene”refer to the unsaturated forms of “alkylene” having double or triplebonds, respectively.

As used herein, a wavy line, “

”, that intersects a single, double or triple bond in any chemicalstructure depicted herein, represent the point attachment of the single,double, or triple bond to the remainder of the molecule.

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. Additionally, for dialkylaminogroups, the alkyl portions can be the same or different and can also becombined to form a 3-7 membered ring with the nitrogen atom to whicheach is attached. Accordingly, a group represented as dialkylamino or—NR^(a)R^(b) is meant to include piperidinyl, pyrrolidinyl, morpholinyl,azetidinyl and the like.

The term “di-(C₁₋₄ alkyl)amino-C₁₋₄ alkyl” refers to an amino groupbearing two C₁₋₄ alkyl groups that can be the same or different (e.g.,methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl andtert-butyl) and which is attached to the remainder of the moleculethrough a C₁₋₄ alkyl group (a one to four carbon alkylene linkinggroup). Examples of di-(C₁₋₄ alkyl)amino-C₁₋₄ alkyl groups includedimethylaminomethyl, 2-(ethyl(methyl)amino)ethyl,3-(dimethylamino)butyl, and the like.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“C₁₋₄ haloalkyl” is mean to include trifluoromethyl,2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,typically aromatic, hydrocarbon group which can be a single ring ormultiple rings (up to three rings) which are fused together or linkedcovalently. The term “heteroaryl” refers to aryl groups (or rings) thatcontain from one to five heteroatoms selected from N, O, and S, whereinthe nitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. A heteroaryl group can be attachedto the remainder of the molecule through a heteroatom. Non-limitingexamples of aryl groups include phenyl, naphthyl and biphenyl, whilenon-limiting examples of heteroaryl groups include pyridyl, pyridazinyl,pyrazinyl, pyrimindinyl, triazinyl, quinolinyl, quinoxalinyl,quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl,benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl,isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl,thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines,benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl,isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl,triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl,thiazolyl, furyl, thienyl and the like. Substituents for each of theabove noted aryl and heteroaryl ring systems are selected from the groupof acceptable substituents described below.

The term “arylalkyl” is meant to include those radicals in which an arylgroup is attached to an alkyl group (e.g., benzyl, phenethyl, and thelike). Similarly, the term “heteroaryl-alkyl” is meant to include thoseradicals in which a heteroaryl group is attached to an alkyl group(e.g., pyridylmethyl, thiazolylethyl, and the like).

The above terms (e.g., “alkyl,” “aryl” and “heteroaryl”), in someembodiments, will refer to both substituted and unsubstituted forms ofthe indicated radical. Preferred substituents for each type of radicalare provided below.

Substituents for the alkyl radicals (including those groups oftenreferred to as alkylene, alkenyl, alkynyl and cycloalkyl) can be avariety of groups selected from: -halogen, —OR′, —NR′R″, —SR′,—SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR′S(O)₂R″, —CN and—NO₂ in a number ranging from zero to (2 m′+1), where m′ is the totalnumber of carbon atoms in such radical. R′, R″ and R′″ eachindependently refer to hydrogen, unsubstituted C₁₋₈ alkyl, unsubstitutedaryl, aryl substituted with 1-3 halogens, unsubstituted C₁₋₈ alkyl, C₁₋₈alkoxy or C₁₋₈ thioalkoxy groups, or unsubstituted aryl-C₁₋₄ alkylgroups. When R′ and R″ are attached to the same nitrogen atom, they canbe combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or7-membered ring. For example, —NR′R″ is meant to include 1-pyrrolidinyland 4-morpholinyl.

Similarly, substituents for the aryl and heteroaryl groups are variedand are generally selected from: -halogen, —OR′, —OC(O)R′, —NR′R″, —SR′,—R′, —CN, —NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′,—NR″C(O)₂R′, —NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR′S(O)₂R″, —N₃,perfluoro(C₁-C₄)alkoxy, and perfluoro(C₁-C₄)alkyl, in a number rangingfrom zero to the total number of open valences on the aromatic ringsystem; and where R′, R″ and R′″ are independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl,C₂₋₈ alkynyl, unsubstituted aryl and heteroaryl, (unsubstitutedaryl)-C₁₋₄ alkyl, and unsubstituted aryloxy-C₁₋₄ alkyl. Other suitablesubstituents include each of the above aryl substituents attached to aring atom by an alkylene tether of from 1-4 carbon atoms.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula-A-(CH₂)_(r)—B—, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituent R′ in —NR′— and —S(O)₂NR′— is selected from hydrogen orunsubstituted C₁₋₆ alkyl.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si).

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of salts derived frompharmaceutically-acceptable inorganic bases include aluminum, ammonium,calcium, copper, ferric, ferrous, lithium, magnesium, manganic,manganous, potassium, sodium, zinc and the like. Salts derived frompharmaceutically-acceptable organic bases include salts of primary,secondary and tertiary amines, including substituted amines, cyclicamines, naturally-occurring amines and the like, such as arginine,betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, malonic, benzoic, succinic, suberic,fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, S. M., et al, “Pharmaceutical Salts”, Journal of PharmaceuticalScience, 1977, 66, 1-19). Certain specific compounds of the presentinvention contain both basic and acidic functionalities that allow thecompounds to be converted into either base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers, regioisomers and individual isomers (e.g., separateenantiomers) are all intended to be encompassed within the scope of thepresent invention. The compounds of the present invention may alsocontain unnatural proportions of atomic isotopes at one or more of theatoms that constitute such compounds. Unnatural proportions of anisotope may be defined as ranging from the amount found in nature to anamount consisting of 100% of the atom in question. For example, thecompounds may incorporate radioactive isotopes, such as for exampletritium (³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C), or non-radioactiveisotopes, such as deuterium (²H) or carbon-13 (¹³C). Such isotopicvariations can provide additional utilities to those described elsewherewith this application. For instance, isotopic variants of the compoundsof the invention may find additional utility, including but not limitedto, as diagnostic and/or imaging reagents, or as cytotoxic/radiotoxictherapeutic agents. Additionally, isotopic variants of the compounds ofthe invention can have altered pharmacokinetic and pharmacodynamiccharacteristics which can contribute to enhanced safety, tolerability orefficacy during treatment. All isotopic variations of the compounds ofthe present invention, whether radioactive or not, are intended to beencompassed within the scope of the present invention.

Compounds of the invention having formula I can exist in differentisomeric forms. As used herein, the terms cis or trans are used in theirconventional sense in the chemical arts, i.e., referring to the positionof the substituents to one another relative to a reference plane, e.g.,a double bond, or a ring system, such as a decalin-type ring system or ahydroquinolone ring system: in the cis isomer, the substituents are onthe same side of the reference plane, in the trans isomer thesubstituents are on opposite sides. Additionally, different conformersare contemplated by the present invention, as well as distinct rotamers.Conformers are conformational isomers that can differ by rotations aboutone or more a bonds. Rotamers are conformers that differ by rotationabout only a single a bond.

II. General

The present invention derives from the discovery that compounds offormula I act as potent antagonists of the CCR1 receptor. The compoundshave in vivo anti-inflammatory activity and have superiorpharmacokinetic properties. Accordingly, the compounds provided hereinare useful in pharmaceutical compositions, methods for the treatment ofCCR1-mediated diseases, and as controls in assays for the identificationof competitive CCR1 antagonists.

III. Compounds

In one aspect, the present invention provides for a compound of FormulaI:

or pharmaceutically acceptable salt, hydrate, solvate, N-oxide orrotamer thereof. In Formula I, each A is independently selected from thegroup consisting of N and CH;

-   X and Z are each independently selected from the group consisting    -   (i) monocyclic or fused-bicyclic aryl and heteroaryl, wherein        the heteroaryl group has from 1-4 heteroatoms as ring members        selected from N, O and S;    -   (ii) monocyclic four-, five-, six- or seven-membered ring        selected from the group consisting of cycloalkane, and        heterocycloalkane, wherein the heterocycloalkane rings have from        1-3 heteroatoms as ring members selected from N, O and S;    -   wherein each of the rings in (i) and (ii) are optionally        substituted with from 1 to 5 substituents selected from halogen,        CN, C₁₋₈ alkyl, C₃₋₈ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,        C₁₋₈ haloalkyl, C₁₋₈ hydroxyalkyl, —OR^(a), —CO₂R^(a),        —SO₂R^(a), —NR^(a)R^(b), —CONR^(a)R^(b), aryl, 5- or 6-membered        heteroaryl, and 3-, 4-, 5- or 6-membered heterocycloalkane        wherein the heteroatoms present as ring vertices of the        heteroaryl and heterocycloalkane rings are selected from N, O        and S, and wherein the alkyl, cycloalkyl, aryl, heteroaryl and        hetereocycloalkane portions of the substituents are optionally        further substituted with 1-3 R^(a): and optionally, two        substituents on adjacent ring vertices are connected to form an        additional 5- or 6-membered ring which is saturated, unsaturated        or aromatic having ring vertices selected from C, O, N and S;    -   R³ is a member selected from the group consisting of H, halogen,        CN, C₁₋₈ alkyl, C₃₋₈ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,        C₁₋₈ haloalkyl, C₁₋₈ hydroxyalkyl, —OR^(a), —CO₂R^(a),        —NR^(a)R^(b), —CONR^(a)R^(b), aryl, 5- or 6-membered heteroaryl,        and 3-, 4-, 5- or 6-membered heterocyclic wherein the        heteroatoms present as ring vertices of the heteroaryl and        heterocyclic rings are selected from N, O and S, and wherein the        alkyl, cycloalkyl, aryl, heteroaryl and hetereocyclic portions        of R³ are optionally further substituted with 1-3 R^(a);    -   R⁴ is a member selected from the group consisting of H, —OR^(a)        and C₁₋₈ alkyl optionally substituted with —OR³ or —NR^(a)R^(b);        or R⁴ is combined with X to form a bicyclic fused ring system;        and        each R^(a) and R^(b) is independently selected from the group        consisting of hydrogen, hydroxyl, halogen, cyano, C₁₋₈ alkyl,        C₁₋₈ alkoxy, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₃₋₆        cycloalkylalkyl, amino, C₁₋₈ alkylamino, di C₁₋₈ alkylamino,        carboxamide, carboxy C₁₋₄ alkyl ester, carboxylic acid, and        —SO₂—C₁₋₈ alkyl.

One of skill in the art will appreciate that substituent recitationsonly refer to those that are generally stable (e.g., less than 20%degradation on storage), such that the group —OR^(a) is not meant toinclude those components wherein R^(a) is alkoxy (which would furnish aperoxy or —OO— alkyl group).

In some selected embodiments, the compounds of Formula I are representedby Formula Ia:

wherein A¹ is N or C(R⁵); A² is N or C(R⁷); and R⁵, R⁶, R⁷ and R⁸ areeach independently selected from H, halogen, CN, C₁₋₈ alkyl, C₃₋₈cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₁₋₈ haloalkyl, C₁₋₈hydroxyalkyl, —OR^(a), —CO₂R^(a), —NR^(a)R^(b), —CONR^(a)R^(b), aryl, 5-or 6-membered heteroaryl, and 3-, 4-, 5- or 6-membered heterocycloalkanewherein the heteroatoms present as ring vertices of the heteroaryl andheterocycloalkane rings are selected from N, O and S, and wherein thealkyl, cycloalkyl, aryl, heteroaryl and hetereocycloalkane portions ofR⁵, R⁶, R⁷ and R⁸ are optionally further substituted with 1-3 R^(a); andoptionally, and optionally, R⁴ and R⁵, R⁴ and R⁸, or adjacent members ofR⁵, R⁶, R⁷ and R⁸ are connected to form an additional 5- or 6-memberedring which is saturated, unsaturated or aromatic having ring verticesselected from C, O, N and S; or a pharmaceutically acceptable salt,hydrate, solvate, rotamer or N-oxide thereof.

In other selected embodiments, the compounds of Formula Ia are thosewherein R⁸ is other than H.

In other selected embodiments, the compounds of Formula 1a arerepresented by Formula Ib:

wherein R¹ and R² are each independently selected from H, halogen, CN,C₁₋₈ alkyl, C₃₋₈ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₁₋₈ haloalkyl,C₁₋₈ hydroxyalkyl, —OR^(a), —CO₂R^(a), —SO₂R^(a), —NR^(a)R^(b),—CONR^(a)R^(b), and 3-, 4-, 5- or 6-membered heterocycloalkane whereinthe heteroatoms present as ring vertices of the heterocycloalkane ringare selected from N, O and S, and wherein the alkyl, cycloalkyl andhetereocycloalkane portions of R¹ and R² are optionally furthersubstituted with 1-3 R^(a).

In selected embodiments of Formula Ib, each R¹ and R² is independentlyselected from H, halogen, CN, C₁₋₈ alkyl, C₁₋₈ haloalkyl, —CO₂R^(a) and—SO₂R^(a).

In other selected embodiments for the compounds of Formula Ib, thecompounds are represented by the structure:

In other selected embodiments for the compounds of Formula Ib and Ib1,the ring portion having N, A¹ and A² as ring vertices is selected from:

In still other selected embodiments for the compounds of Formula Ib andIb1, the ring portion having N, A¹ and A² as ring vertices is selectedfrom:

wherein R⁷ is H or Cl, and R⁸ is C₁₋₈ alkyl optionally substituted with1 or 2 R^(a).

In still other selected embodiments of Formula Ib or Ib1, R⁴ is H orCH₃.

Returning to Formula I, some selected embodiments are those compoundsrepresented by Formula Ib2:

wherein R¹ is Cl or F; R³ is selected from the group consisting of C₁₋₈alkyl, C₁₋₈ haloalkyl, C₁₋₈ hydroxyalkyl, wherein the alkyl portions ofR³ are optionally further substituted with 1-3 R³; and wherein R⁷ and R⁸are not joined to form a ring

In still other selected embodiments, the compounds of Formula I, Ib, Ib1and Ib2 are represented by Formulae Ib2a, Ib2b and Ib2c.

In some selected embodiments, the compounds are represented by FormulaIc:

wherein the subscript n is 0 or 1.

In some selected embodiments, the compounds are represented by FormulaIb3:

In some selected embodiments of Formula Ib, the compounds arerepresented by Formulae Ib2d, Ib2e and Ib2f.

In selected embodiments of any of Formulae I, Ia, Ib, Ib1, Ib2, Ib2a,Ib2b, Ib2c, Ib2d, Ib2e, Ib2f, Ib3 and Ic, R³ is C₁₋₈ alkyl.

Preparation of Compounds

The schemes in the Examples below provide certain synthetic routes thatcan be followed to access certain compounds of the present invention.Other routes or modification of the routes presented below would bereadily apparent to a skilled artisan and are within the scope of thepresent invention.

IV. Pharmaceutical Compositions

In addition the compounds provided above, the compositions formodulating CCR1 activity in humans and animals will typically contain apharmaceutical carrier or diluent.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacyand drug delivery. All methods include the step of bringing the activeingredient into association with the carrier which constitutes one ormore accessory ingredients. In general, the pharmaceutical compositionsare prepared by uniformly and intimately bringing the active ingredientinto association with a liquid carrier or a finely divided solid carrieror both, and then, if necessary, shaping the product into the desiredformulation. In the pharmaceutical composition the active objectcompound is included in an amount sufficient to produce the desiredeffect upon the process or condition of diseases.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions and self emulsifications as described in U.S. Pat. No.6,451,339, hard or soft capsules, syrups, elixirs, solutions, buccalpatch, oral gel, chewing gum, chewable tablets, effervescent powder andeffervescent tablets. Compositions intended for oral use may be preparedaccording to any method known to the art for the manufacture ofpharmaceutical compositions and such compositions may contain one ormore agents selected from the group consisting of sweetening agents,flavoring agents, coloring agents, antioxidants and preserving agents inorder to provide pharmaceutically elegant and palatable preparations.Tablets contain the active ingredient in admixture with non-toxicpharmaceutically acceptable excipients which are suitable for themanufacture of tablets. These excipients may be for example, inertdiluents, such as cellulose, silicon dioxide, aluminum oxide, calciumcarbonate, sodium carbonate, glucose, mannitol, sorbitol, lactose,calcium phosphate or sodium phosphate; granulating and disintegratingagents, for example, corn starch, or alginic acid; binding agents, forexample PVP, cellulose, PEG, starch, gelatin or acacia, and lubricatingagents, for example magnesium stearate, stearic acid or talc. Thetablets may be uncoated or they may be coated, enterically or otherwise,by known techniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate may be employed. They may also becoated by the techniques described in the U.S. Pat. Nos. 4,256,108;4,166,452; and 4,265,874 to form osmotic therapeutic tablets for controlrelease.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.Additionally, emulsions can be prepared with a non-water miscibleingredient such as oils and stabilized with surfactants such asmono-diglycerides, PEG esters and the like.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxy-ethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents. Oral solutions can be prepared in combination with, for example,cyclodextrin, PEG and surfactants.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials include cocoa butter andpolyethylene glycols. Additionally, the compounds can be administeredvia ocular delivery by means of solutions or ointments. Still further,transdermal delivery of the subject compounds can be accomplished bymeans of iontophoretic patches and the like. For topical use, creams,ointments, jellies, solutions or suspensions, etc., containing thecompounds of the present invention are employed. As used herein, topicalapplication is also meant to include the use of mouth washes andgargles.

The compounds of the invention may be formulated for depositing into amedical device, which may include any of variety of conventional grafts,stents, including stent grafts, catheters, balloons, baskets or otherdevice that can be deployed or permanently implanted within a bodylumen. As a particular example, it would be desirable to have devicesand methods which can deliver compounds of the invention to the regionof a body which has been treated by interventional technique.

In exemplary embodiment, the inhibitory agent of this invention may bedeposited within a medical device, such as a stent, and delivered to thetreatment site for treatment of a portion of the body.

Stents have been used as delivery vehicles for therapeutic agents (i.e.,drugs). Intravascular stents are generally permanently implanted incoronary or peripheral vessels. Stent designs include those of U.S. Pat.No. 4,733,655 (Palmaz), U.S. Pat. No. 4,800,882 (Gianturco), or U.S.Pat. No. 4,886,062 (Wiktor). Such designs include both metal andpolymeric stents, as well as self-expanding and balloon-expandablestents. Stents may also used to deliver a drug at the site of contactwith the vasculature, as disclosed in U.S. Pat. No. 5,102,417 (Palmaz)and in International Patent Application Nos. WO 91/12779 (Medtronic,Inc.) and WO 90/13332 (Cedars-Sanai Medical Center), U.S. Pat. No.5,419,760 (Narciso, Jr.) and U.S. Pat. No. 5,429,634 (Narciso, Jr.), forexample. Stents have also been used to deliver viruses to the wall of alumen for gene delivery, as disclosed in U.S. Pat. No. 5,833,651(Donovan et al.).

The term “deposited” means that the inhibitory agent is coated,adsorbed, placed, or otherwise incorporated into the device by methodsknown in the art. For example, the inhibitory agent may be embedded andreleased from within (“matrix type”) or surrounded by and releasedthrough (“reservoir type”) polymer materials that coat or span themedical device. In the later example, the inhibitory agent may beentrapped within the polymer materials or coupled to the polymermaterials using one or more the techniques for generating such materialsknown in the art. In other formulations, the inhibitory agent may belinked to the surface of the medical device without the need for acoating by means of detachable bonds and release with time, can beremoved by active mechanical or chemical processes, or are in apermanently immobilized form that presents the inhibitory agent at theimplantation site.

In one embodiment, the inhibitory agent may be incorporated with polymercompositions during the formation of biocompatible coatings for medicaldevices, such as stents. The coatings produced from these components aretypically homogeneous and are useful for coating a number of devicesdesigned for implantation.

The polymer may be either a biostable or a bioabsorbable polymerdepending on the desired rate of release or the desired degree ofpolymer stability, but a bioabsorbable polymer is preferred for thisembodiment since, unlike a biostable polymer, it will not be presentlong after implantation to cause any adverse, chronic local response.Bioabsorbable polymers that could be used include, but are not limitedto, poly(L-lactic acid), polycaprolactone, polyglycolide (PGA),poly(lactide-co-glycolide) (PLLA/PGA), poly(hydroxybutyrate),poly(hydroxybutyrate-co-valerate), polydioxanone, polyorthoester,polyanhydride, poly(glycolic acid), poly(D-lactic acid), poly(L-lacticacid), poly(D,L-lactic acid), poly(D,L-lactide) (PLA), poly (L-lactide)(PLLA), poly(glycolic acid-co-trimethylene carbonate) (PGA/PTMC),polyethylene oxide (PEO), polydioxanone (PDS), polyphosphoester,polyphosphoester urethane, poly(amino acids), cyanoacrylates,poly(trimethylene carbonate), poly(iminocarbonate), copoly(ether-esters)(e.g., PEO/PLA), polyalkylene oxalates, polyphosphazenes andbiomolecules such as fibrin, fibrinogen, cellulose, starch, collagen andhyaluronic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates,cross linked or amphipathic block copolymers of hydrogels, and othersuitable bioabsorbable poplymers known in the art. Also, biostablepolymers with a relatively low chronic tissue response such aspolyurethanes, silicones, and polyesters could be used and otherpolymers could also be used if they can be dissolved and cured orpolymerized on the medical device such as polyolefins, polyisobutyleneand ethylene-alphaolefin copolymers; acrylic polymers and copolymers,vinyl halide polymers and copolymers, such as polyvinyl chloride;polyvinylpyrrolidone; polyvinyl ethers, such as polyvinyl methyl ether;polyvinylidene halides, such as polyvinylidene fluoride andpolyvinylidene chloride; polyacrylonitrile, polyvinyl ketones; polyvinylaromatics, such as polystyrene, polyvinyl esters, such as polyvinylacetate; copolymers of vinyl monomers with each other and olefins, suchas ethylene-methyl methacrylate copolymers, acrylonitrile-styrenecopolymers, ABS resins, and ethylene-vinyl acetate copolymers; pyrancopolymer; polyhydroxy-propyl-methacrylamide-phenol;polyhydroxyethyl-aspartamide-phenol; polyethyleneoxide-polylysinesubstituted with palmitoyl residues; polyamides, such as Nylon 66 andpolycaprolactam; alkyd resins, polycarbonates; polyoxymethylenes;polyimides; polyethers; epoxy resins, polyurethanes; rayon;rayon-triacetate; cellulose, cellulose acetate, cellulose butyrate;cellulose acetate butyrate; cellophane; cellulose nitrate; cellulosepropionate; cellulose ethers; and carboxymethyl cellulose.

Polymers and semipermeable polymer matrices may be formed into shapedarticles, such as valves, stents, tubing, prostheses and the like.

In one embodiment of the invention, the inhibitory agent of theinvention is coupled to a polymer or semipermeable polymer matrix thatis formed as a stent or stent-graft device.

Typically, polymers are applied to the surface of an implantable deviceby spin coating, dipping or spraying. Additional methods known in theart can also be utilized for this purpose. Methods of spraying includetraditional methods as well as microdeposition techniques with an inkjettype of dispenser. Additionally, a polymer can be deposited on animplantable device using photo-patterning to place the polymer on onlyspecific portions of the device. This coating of the device provides auniform layer around the device which allows for improved diffusion ofvarious analytes through the device coating.

In preferred embodiments of the invention, the inhibitory agent isformulated for release from the polymer coating into the environment inwhich the medical device is placed. Preferably, the inhibitory agent isreleased in a controlled manner over an extended time frame (e.g.,months) using at least one of several well-known techniques involvingpolymer carriers or layers to control elution. Some of these techniqueswere previously described in U.S. Patent Application 20040243225A1.

Moreover, as described for example in U.S. Pat. No. 6,770,729, thereagents and reaction conditions of the polymer compositions can bemanipulated so that the release of the inhibitory agent from the polymercoating can be controlled. For example, the diffusion coefficient of theone or more polymer coatings can be modulated to control the release ofthe inhibitory agent from the polymer coating. In a variation on thistheme, the diffusion coefficient of the one or more polymer coatings canbe controlled to modulate the ability of an analyte that is present inthe environment in which the medical device is placed (e.g. an analytethat facilitates the breakdown or hydrolysis of some portion of thepolymer) to access one or more components within the polymer composition(and for example, thereby modulate the release of the inhibitory agentfrom the polymer coating). Yet another embodiment of the inventionincludes a device having a plurality of polymer coatings, each having aplurality of diffusion coefficients. In such embodiments of theinvention, the release of the inhibitory agent from the polymer coatingcan be modulated by the plurality of polymer coatings.

In yet another embodiment of the invention, the release of theinhibitory agent from the polymer coating is controlled by modulatingone or more of the properties of the polymer composition, such as thepresence of one or more endogenous or exogenous compounds, oralternatively, the pH of the polymer composition. For example, certainpolymer compositions can be designed to release a inhibitory agent inresponse to a decrease in the pH of the polymer composition.Alternatively, certain polymer compositions can be designed to releasethe inhibitory agent in response to the presence of hydrogen peroxide.

III. Methods of Treating Diseases Modulated by CCR1

In yet another aspect, the present invention provides methods oftreating CCR1-mediated conditions or diseases by administering to asubject having such a disease or condition, a therapeutically effectiveamount of a compound of formula I above. The “subject” is defined hereinto include animals such as mammals, including, but not limited to,primates (e.g., humans), cows, sheep, goats, horses, dogs, cats,rabbits, rats, mice and the like.

CCR1 provides a target for interfering with or promoting specificaspects of immune cell functions, or more generally, with functionsassociated with CCR1 expression on a wide range of cell types in amammal, such as a human. Compounds that inhibit CCR1, are particularlyuseful for modulating monocyte, macrophage, lymphocyte, granulocyte, NKcell, mast cells, dendritic cell, and certain immune derived cell (forexample, osteoclasts) function for therapeutic purposes. Accordingly,the present invention is directed to compounds which are useful in theprevention and/or treatment of a wide variety of inflammatory andimmunoregulatory disorders and diseases (see Saeki, et al., CurrentPharmaceutical Design 9:1201-1208 (2003)).

For example, an instant compound that inhibits one or more functions ofCCR1 may be administered to inhibit (i.e., reduce or prevent)inflammation or cellular infiltration associated with an immunedisorder. As a result, one or more inflammatory processes, such asleukocyte emigration or infiltration, chemotaxis, exocytosis (e.g., ofenzymes, histamine) or inflammatory mediator release, can be inhibited.For example, monocyte infiltration to an inflammatory site (e.g., anaffected joint in arthritis, or into the CNS in MS) can be inhibitedaccording to the present method.

Similarly, an instant compound that promotes one or more functions ofCCR1 is administered to stimulate (induce or enhance) an inflammatoryresponse, such as leukocyte emigration, chemotaxis, exocytosis (e.g., ofenzymes, histamine) or inflammatory mediator release, resulting in thebeneficial stimulation of inflammatory processes. For example, monocytescan be recruited to combat bacterial infections.

Diseases and conditions associated with inflammation, immune disordersand infection can be treated using the method of the present invention.In a preferred embodiment, the disease or condition is one in which theactions of immune cells such monocyte, macrophage, lymphocyte,granulocyte, NK cell, mast cell, dendritic cell, or certain immunederived cell (for example, osteoclasts) are to be inhibited or promoted,in order to modulate the inflammatory or autoimmune response.

In one group of embodiments, diseases or conditions, including chronicdiseases, of humans or other species can treated with modulators of CCR1function. These diseases or conditions include: (1) allergic diseasessuch as systemic anaphylaxis or hypersensitivity responses, drugallergies, insect sting allergies and food allergies, (2) inflammatorybowel diseases, such as Crohn's disease, ulcerative colitis, ileitis andenteritis, (3) vaginitis, (4) psoriasis and inflammatory dermatoses suchas dermatitis, eczema, atopic dermatitis, allergic contact dermatitis,urticaria and pruritus, (5) vasculitis, (6) spondyloarthropathies, (7)scleroderma, (8) asthma and respiratory allergic diseases such asasthma, allergic asthma, allergic rhinitis, hypersensitivity lungdiseases and the like, (9) autoimmune diseases, such as fibromyalagia,scleroderma, ankylosing spondylitis, juvenile RA, Still's disease,polyarticular juvenile RA, pauciarticular juvenile RA, polymyalgiarheumatica, Takuyasu arthritis, rheumatoid arthritis, psoriaticarthritis, osteoarthritis, polyarticular arthritis, multiple sclerosis,systemic lupus erythematosus, type I diabetes, type II diabetes, type Idiabetes (recent onset), optic neuritis, glomerulonephritis, and thelike, (10) graft rejection including allograft rejection and acute andchronic graft-vs-host disease, (11) fibrosis (e.g. pulmonary fibrosis(i.e. idiopathic pulmonary fibrosis, interstitial pulmonary fibrosis),fibrosis associated with end-stage renal disease, fibrosis caused byradiation, tubulointerstitial fibrosis, subepithelieal fibrosis,scleroderma (progressive systemic sclerosis), hepatic fibrosis(including that caused by alcoholic or viral hepatitis), primary andsecondary cirrhosis), (12) acute and chronic lung inflammation (chronicobstructive pulmonary disease, chronic bronchitis, adult respiratorydistress syndrome, respiratory distress syndrome of infancy, immunecomplex alveolitis) and (13) other diseases in which undesiredinflammatory responses or immune disorders are to be inhibited, such ascardiovascular disease including atherosclerosis, vascular inflammationresulting from tissue transplant or during restenosis (including, butnot limited to restenosis following angioplasty and/or stent insertion),other acute and chronic inflammatory conditions such as myositis,neurodegenerative diseases (e.g., Alzheimer's disease), encephalitis,meningitis, hepatitis, nephritis, sepsis, sarcoidosis, allergicconjunctivitis, otitis, sinusitis, synovial inflammation caused byarthroscopy, hyperuremia, trauma, ischaemia reperfusion injury, nasalpolyosis, preeclampsia, oral lichen planus, Guillina-Barre syndrome,granulomatous diseases, conditions associated with leptin production,Behcet's syndrome and gout and in wound healing applications (14) immunemediated food allergies such as Celiac disease (15) diseases ofosteoclast dysregulation including osteoporosis and osteolytic bonediseases associated with cancers such as multiple myeloma.

In another group of embodiments, diseases or conditions can be treatedwith modulators of CCR1 function. Examples of diseases to be treatedwith modulators of CCR1 function include cancers (both primary andmetastatic) (e.g., multiple myeloma; Hata, H., Leukemia & Lymphoma,2005, 46(7); 967-972), cardiovascular diseases, diseases in whichangiogenesis or neovascularization play a role (neoplastic diseases,retinopathy and macular degeneration), infectious diseases (viralinfections, e.g., HIV infection, and bacterial infections) andimmunosuppressive diseases such as organ transplant conditions and skintransplant conditions. The term “organ transplant conditions” is meantto include bone marrow transplant conditions and solid organ (e.g.,kidney, liver, lung, heart, pancreas or combination thereof) transplantconditions.

Pharmaceutical compositions of this invention can also inhibit theproduction of metalloproteinases and cytokines at inflammatory sites,either directly or indirectly (as a consequence of decreasing cellinfiltration) thus providing benefit for diseases or conditions linkedto these cytokines.

The compounds of the present invention are accordingly useful in theprevention and treatment of a wide variety of inflammatory andimmunoregulatory disorders and diseases.

Depending on the disease to be treated and the subject's condition, thecompounds of the present invention may be administered by oral,parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV,intracisternal injection or infusion, subcutaneous injection, orimplant), by inhalation spray, nasal, vaginal, rectal, sublingual, ortopical routes of administration and may be formulated, alone ortogether, in suitable dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carriers, adjuvants and vehiclesappropriate for each route of administration.

Those of skill in the art will understand that agents that modulate CCR1activity can be combined in treatment regimens with other therapeuticagents and/or with chemotherapeutic agents or radiation. In some cases,the amount of chemotherapeutic agent or radiation is an amount whichwould be sub-therapeutic if provided without combination with acomposition of the invention. Those of skill in the art will appreciatethat “combinations” can involve combinations in treatments (i.e., two ormore drugs can be administered as a mixture, or at least concurrently orat least introduced into a subject at different times but such that bothare in the bloodstream of a subject at the same time). Additionally,compositions of the current invention may be administered prior to orsubsequent to a second therapeutic regimen, for instance prior to orsubsequent to a dose of chemotherapy or irradiation.

In the treatment or prevention of conditions which require chemokinereceptor modulation an appropriate dosage level will generally be about0.001 to 100 mg per kg patient body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.01 to about 25 mg/kg per day; more preferably about 0.05to about 10 mg/kg per day. A suitable dosage level may be about 0.01 to25 mg/kg per day, about 0.05 to 10 mg/kg per day, or about 0.1 to 5mg/kg per day. Within this range the dosage may be 0.005 to 0.05, 0.05to 0.5 or 0.5 to 5.0 mg/kg per day. For oral administration, thecompositions are preferably provided in the form of tablets containing1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0,10.0, 15.0. 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0,400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of theactive ingredient for the symptomatic adjustment of the dosage to thepatient to be treated. The compounds may be administered on a regimen of1 to 4 times per day, preferably once or twice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, hereditary characteristics, generalhealth, sex and diet of the subject, as well as the mode and time ofadministration, rate of excretion, drug combination, and the severity ofthe particular condition for the subject undergoing therapy.

Diseases and conditions associated with inflammation, immune disorder,infection and cancer can be treated or prevented with the presentcompounds, compositions, and methods.

The compounds and compositions of the present invention can be combinedwith other compounds and compositions having related utilities toprevent and treat the condition or disease of interest, such asinflammatory or autoimmune disorders, conditions and diseases, includinginflammatory bowel disease, rheumatoid arthritis, osteoarthritis,psoriatic arthritis, polyarticular arthritis, multiple sclerosis,allergic diseases, psoriasis, atopic dermatitis and asthma, and thosepathologies noted above.

For example, in the treatment or prevention of inflammation orautimmunity or for example arthritis associated bone loss, the presentcompounds and compositions may be used in conjunction with ananti-inflammatory or analgesic agent such as an opiate agonist, alipoxygenase inhibitor, such as an inhibitor of 5-lipoxygenase, acyclooxygenase inhibitor, such as a cyclooxygenase-2 inhibitor, aninterleukin inhibitor, such as an interleukin-1 inhibitor, an NMDAantagonist, an inhibitor of nitric oxide or an inhibitor of thesynthesis of nitric oxide, a non steroidal anti-inflammatory agent, or acytokine-suppressing anti-inflammatory agent, for example with acompound such as acetaminophen, aspirin, codeine, fentanyl, ibuprofen,indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, asteroidal analgesic, sufentanyl, sunlindac, tenidap, and the like.Similarly, the instant compounds and compositions may be administeredwith an analgesic listed above; a potentiator such as caffeine, an H2antagonist (e.g., ranitidine), simethicone, aluminum or magnesiumhydroxide; a decongestant such as phenylephrine, phenylpropanolamine,pseudoephedrine, oxymetazoline, ephinephrine, naphazoline,xylometazoline, propylhexedrine, or levo desoxy ephedrine; anantitussive such as codeine, hydrocodone, caramiphen, carbetapentane, ordextromethorphan; a diuretic; and a sedating or non sedatingantihistamine.

Likewise, compounds and compositions of the present invention may beused in combination with other drugs that are used in the treatment,prevention, suppression or amelioration of the diseases or conditionsfor which compounds and compositions of the present invention areuseful. Such other drugs may be administered, by a route and in anamount commonly used therefor, contemporaneously or sequentially with acompound or composition of the present invention. When a compound orcomposition of the present invention is used contemporaneously with oneor more other drugs, a pharmaceutical composition containing such otherdrugs in addition to the compound or composition of the presentinvention is preferred. Accordingly, the pharmaceutical compositions ofthe present invention include those that also contain one or more otheractive ingredients or therapeutic agents, in addition to a compound orcomposition of the present invention. Examples of other therapeuticagents that may be combined with a compound or composition of thepresent invention, either administered separately or in the samepharmaceutical compositions, include, but are not limited to: (a) VLA-4antagonists, (b) corticosteroids, such as beclomethasone,methylprednisolone, betamethasone, prednisone, prenisolone,dexamethasone, fluticasone, hydrocortisone, budesonide, triamcinolone,salmeterol, salmeterol, salbutamol, formeterol; (c) immunosuppressantssuch as cyclosporine (cyclosporine A, Sandimmune®, Neoral®), tacrolirnus(FK-506, Prograf®), rapamycin (sirolimus, Rapamune®), Tofacitinib(Xeljanz®) and other FK-506 type immunosuppressants, and rnycophenolate,e.g., mycophenolate mofetil (CellCept®); (d) antihistamines(H1-histamine antagonists) such as bromopheniramine, chlorpheniramine,dexchloipheniramine, triprolidine, clemastine, diphenhydramine,diphenylpyraline, tripelennamine, hydroxyzine, methdilazine,promethazine, trimeprazine, azatadine, cyproheptadine, antazoline,pheniramine pyrilamine, astemizole, terfenadine, loratadine, cetirizine,fexofenadine, descarboethoxyloratadine, and the like; (e) non steroidalanti asthmatics (e.g., terbutaline, metaproterenol, fenoterol,isoetharine, albuterol, bitolterol and pirbuterol), theophylline,cromolyn sodium, atropine, ipratropium bromide, leukotriene antagonists(e.g., zafmlukast, montelukast, pranlukast, iralukast, pobilukast andSKB-106,203), leukotriene biosynthesis inhibitors (zileuton, BAY-1005);(f) non steroidal anti-inflammatory agents (NSAIDs) such as propionicacid derivatives (e.g., alminoprofen, benoxaprofen, bucloxic acid,carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen,indoprofen, ketoprofen, niroprofen, naproxen, oxaprozin, pirprofen,pranoprofen, suprofen, tiaprofenic acid and tioxaprofen), acetic acidderivatives (e.g., indomethacin, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin andzomepirac), fenamic acid derivatives (e.g., flufenamic acid,meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid),biphenylcarboxylic acid derivatives (e.g., diflunisal and flufenisal),oxicams (e.g., isoxicam, piroxicam, sudoxicam and tenoxican),salicylates (e.g., acetyl salicylic acid and sulfasalazine) and thepyrazolones (e.g., apazone, bezpiperylon, feprazone, mofebutazone,oxyphenbutazone and phenylbutazone); (g) cyclooxygenase-2 (COX-2)inhibitors such as celecoxib (Celebrex®) and rofecoxib (Vioxx®); (h)inhibitors of phosphodiesterase type IV (PDE IV); (i) gold compoundssuch as auranofin and aurothioglucose, (j) etanercept (Enbrel®), (k)antibody therapies such as orthoclone (OKT3), daclizumab (Zenapax®),basiliximab (Simulect®) and infliximab (Remicade®), adalimumab(Humira®), golimumab (Simponi®), rituximab (Rituxan®), tocilizumab(Actemra®), (l) other antagonists of the chemokine receptors, especiallyCCR5, CXCR2, CXCR3, CCR2, CCR3, CCR4, CCR7, CX₃CR1 and CXCR6; (m)lubricants or emollients such as petrolatum and lanolin, (n) keratolyticagents (e.g., tazarotene), (o) vitamin D₃ derivatives, e.g.,calcipotriene or calcipotriol (Dovonex®), (p) PUVA, (q) anthralin(Drithrocreme®), (r) etretinate (Tegison®) and isotretinoin and (s)multiple sclerosis therapeutic agents such as interferon β-1β(Betaseron®), interferon (β-1α (Avonex®), azathioprine (Imurek®,Imuran®), glatiramer acetate (Capoxone®), a glucocorticoid (e.g.,prednisolone) and cyclophosphamide (t) DMARDS such as methotrexate andleflunomide (u) other compounds such as 5-aminosalicylic acid andprodrugs thereof; hydroxychloroquine; D-penicillamine; antimetabolitessuch as azathioprine, 6-mercaptopurine and methotrexate; DNA synthesisinhibitors such as hydroxyurea and microtubule disrupters such ascolchicine and proteasome inhibitors such as bortezomib (Velcade®). Theweight ratio of the compound of the present invention to the secondactive ingredient may be varied and will depend upon the effective doseof each ingredient. Generally, an effective dose of each will be used.Thus, for example, when a compound of the present invention is combinedwith an NSAID the weight ratio of the compound of the present inventionto the NSAID will generally range from about 1000:1 to about 1:1000,preferably about 200:1 to about 1:200. Combinations of a compound of thepresent invention and other active ingredients will generally also bewithin the aforementioned range, but in each case, an effective dose ofeach active ingredient should be used.

IV. Examples

The following examples are offered to illustrate, but not to limit theclaimed invention.

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMR wererecorded on a Varian Mercury 400 MHz NMR spectrometer. Significant peaksare provided relative to TMS and are tabulated in the order:multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m,multiplet) and number of protons. Mass spectrometry results are reportedas the ratio of mass over charge, followed by the relative abundance ofeach ion (in parenthesis). In tables, a single m/e value is reported forthe M+H (or, as noted, M−H) ion containing the most common atomicisotopes. Isotope patterns correspond to the expected formula in allcases. Electrospray ionization (ESI) mass spectrometry analysis wasconducted on a Hewlett-Packard MSD electrospray mass spectrometer usingthe HP1100 HPLC equipped with an Agilent Zorbax SB-C18, 2.1×50 mm, 5μcolumn for sample delivery. Normally the analyte was dissolved inmethanol at 0.1 mg/mL and 1 microlitre was infused with the deliverysolvent into the mass spectrometer, which scanned from 100 to 1500daltons. All compounds could be analyzed in the positive ESI mode, usingacetonitrile/water with 1% formic acid as the delivery solvent. Thecompounds provided below could also be analyzed in the negative ESImode, using 2 mM NH₄OAc in acetonitrile/water as delivery system.

The following abbreviations are used in the Examples and throughout thedescription of the invention: HPLC, High Pressure Liquid Chromatography;DMF, Dimethyl formamide; TFA, Trifluoroacetic Acid; THF,Tetrahydrofuran; EtOAc, Ethyl acetate; BOC₂O, di-tertbutyl dicarbonateor BOC anhydride; HPLC, High Pressure Liquid Chromatography; DIPEA,Diisopropyl ethylamine; HBTU,O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate;dppf, 1, 1′-Bis(diphenylphosphino)ferrocene; Pd₂(dba)₃,Tris(dibenzylideneacetone)dipalladium(0); DIPEA, diisopropylethylamine;DMP, dimethylphthalate; Me, methyl; Et, ethyl; DCM, dichloromethane.

Compounds within the scope of this invention can be synthesized asdescribed below, using a variety of reactions known to the skilledartisan. One skilled in the art will also recognize that alternativemethods may be employed to synthesize the target compounds of thisinvention, and that the approaches described within the body of thisdocument are not exhaustive, but do provide broadly applicable andpractical routes to compounds of interest.

Certain molecules claimed in this patent can exist in differentenantiomeric and diastereomeric forms and all such variants of thesecompounds are claimed.

The detailed description of the experimental procedures used tosynthesize key compounds in this text lead to molecules that aredescribed by the physical data identifying them as well as by thestructural depictions associated with them.

Those skilled in the art will also recognize that during standard workup procedures in organic chemistry, acids and bases are frequently used.Salts of the parent compounds are sometimes produced, if they possessthe necessary intrinsic acidity or basicity, during the experimentalprocedures described within this patent.

EXAMPLES Example 1 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-N-[1-(4-fluorophenyl)-5-methylpyrazol-4-yl]acetamide

To a solution of 1-(4-fluorophenyl)-5-isopropylpyrazol-4-amine (0.03 g,0.16 mmol) in DMF (1.0 mL) was added2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]acetic acid (0.038g, 0.16 mmol), followed by diisopropylethylamine (0.042 g, 0.32 mmol)and HATU (0.067 g, 0.18 mmol). After stirring at room temperature for 1h, the reaction was diluted with water and the aqueous layer wasextracted with ethyl acetate (2×5 mL). The combined organic layers werewashed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo.The resulting crude product was purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to give the titlecompound as a white solid (0.02 g, 0.048 mmol, 30%). ¹H NMR (400 MHz,CDCl₃) δ 7.93 (s, 1 H), 7.37 (dd, J=8.9, 4.7 Hz, 1 H), 7.26 (d, J=0.4Hz, 2 H), 7.18 (dd, J=8.9, 8.1 Hz, 2 H), 4.94 (s, 2 H), 2.39 (s, 3 H),2.16 (s, 3 H): MS: (ES) m/z calculated for C₁₇H₁₄ClF₄N₅O [M+H]⁺ 416.1,found 416.0.

Example 2 Synthesis ofN-[1-(4-fluorophenyl)-5-methylpyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

To a solution of 1-(4-fluorophenyl)-5-methylpyrazol-4-amine (0.03 g,0.16 mmol) in DMF (1.0 mL) was added2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanoic acid (0.035 g,0.16 mmol), followed by diisopropylethylamine (0.042 g, 0.32 mmol) andHATU (0.067 g, 0.18 mmol). After stirring at room temperature for 1 h,the reaction was diluted with water, and the aqueous layer was extractedwith ethyl acetate (2×5 mL). The combined organic layers were washedwith brine, dried (Na₂SO₄), filtered, and concentrated in vacuo. Theresulting crude product was purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to give the title compound asa white solid (0.015 g, 0.038 mmol, 24%). ¹H NMR (400 MHz, CDCl₃) δ 8.93(s, 1 H), 7.93 (s, 1 H), 7.81 (d, J=1.3 Hz, 1 H), 7.42-7.32 (m, 2 H),7.24-7.14 (m, 2 H), 5.35 (q, J=6.8 Hz, 1 H), 2.48 (s, 3 H), 2.19 (s, 3H), 1.86 (d, J=7.0 Hz, 3 H); MS: (ES) m % z calculated for C₁₈H₁₇F₄N₅O[M+H]⁺ 396.1, found 396.1.

Example 3 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-N-[5-ethyl-1-(4-fluorophenyl)pyrazol-4-yl]acetamide

To a solution of 5-ethyl-1-(4-fluorophenyl)pyrazol-4-amine (0.02 g, 0.1mmol) in DMF (1.0 mL) was added2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]acetic acid (0.024g, 0.10 mmol), followed by diisopropylethylamine (0.042 g, 0.32 mmol)and HATU (0.045 g, 0.12 mmol). After stirring at room temperature for 1h, the reaction was diluted with water, and the aqueous layer wasextracted with ethyl acetate (2×5 mL). The combined organic layers werewashed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo.The resulting crude product was purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to give the titlecompound as a white solid (0.015 g, 0.035 mmol, 35%). ¹H NMR (400 MHz,CDCl₃) δ 7.97 (s, 1 H), 7.92 (s, 1 H), 7.40-7.31 (m, 2 H), 7.17 (dd,J=8.9, 8.1 Hz, 2 H), 4.93 (s, 2 H), 2.56 (q, J=7.6 Hz, 2 H), 2.39 (s, 3H), 1.01 (t, J=7.6 Hz, 3 H); MS: (ES) m/z calculated for C₁₈H₁₆ClF₄N₅O[M+H]⁺ 430.1, found 430.0.

Example 4 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-N-[1-(4-fluorophenyl)-5-isopropylpyrazol-4-yl]acetamide

To a solution of 1-(4-fluorophenyl)-5-isopropylpyrazol-4-amine (0.035 g,0.16 mmol) in DMF (1.0 mL) was added2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]acetic acid (0.039g, 0.16 mmol), followed by diisopropylethylamine (0.042 g, 0.32 mmol)and HATU (0.073 g, 0.19 mmol). After stirring at room temperature for 1h, the reaction was diluted with water, and the aqueous layer wasextracted with ethyl acetate (2×5 mL). The combined organic layers werewashed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo.The resulting crude product was purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to give the titlecompound as a white solid (0.020 g, 0.045 mmol, 28%). ¹H NMR (400 MHz,CDCl₃) δ 7.99 (t, J=0.5 Hz, 1 H), 7.74 (s, 1 H), 7.35-7.24 (m, 2 H),7.20-7.11 (m, 2 H), 4.92 (s, 2 H), 3.02-2.90 (m, 1 H), 2.39 (s, 3 H), δ1.13 (d, J=7.2 Hz, 6 H); MS: (ES) m/z calculated for C₁₉H₁₈ClF₄N₅O[M+H]⁺ 444.1, found 443.9.

Example 5 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-N-[1-(4-fluorophenyl)-5-phenylpyrazol-4-yl]acetamide

To a solution of 1-(4-fluorophenyl)-5-phenylpyrazol-4-amine (0.03 g,0.12 mmol) in DMF (1.0 mL) was added2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]acetic acid (0.024g, 0.12 mmol), followed by diisopropylethylamine (0.031 g, 0.24 mmol)and HATU (0.055 g, 0.14 mmol). After stirring at room temperature for 1h, the reaction was diluted with water and the aqueous layer wasextracted with ethyl acetate (2×5 mL). The combined organic layers werewashed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo.The resulting crude product was purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to give the titlecompound as a white solid (0.020 g, 0.045 mmol, 28%). ¹H NMR (400 MHz,CDCl₃) δ 8.29 (s, 1 H), 7.55 (s, 1 H), 7.44-7.34 (m, 3 H), 7.22-7.14 (m,2 H), 7.07-6.93 (m, 4 H), 4.86 (s, 2 H), 2.29 (s, 3H); MS: (ES) m/zcalculated for C₂₂H₁₆ClF₄N₅O [M+H]⁺ 478.1, found 477.8.

Example 6 Synthesis ofN-[1-(4-fluorophenyl)-5-isopropylpyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]butanamide

To a solution of 1-(4-fluorophenyl)-5-isopropylpyrazol-4-amine (0.03 g,0.14 mmol) in DMF (1.0 mL) was added2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]butanoic acid (0.032 g,0.14 mmol), followed by diisopropylethylamine (0.042 g, 0.32 mmol) andHATU (0.057 g, 0.15 mmol). After stirring at room temperature for 1 h,the reaction was diluted with water and the aqueous layer was extractedwith ethyl acetate (2×5 mL). The combined organic layers were washedwith brine, dried (Na₂SO₄), filtered, and concentrated in vacuo. Theresulting crude product was purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to give the title compound asa white solid (0.015 g, 0.034 mmol, 25%). ¹H NMR (400 MHz, CDCl₃) δ 7.85(s, 1 H), 7.47 (q, J=1.2 Hz, 1 H), 7.34-7.24 (m, 2 H), 7.21-7.11 (m, 2H), 6.98 (s, 1 H), 4.65 (dd, J=10.1, 5.3 Hz, 1 H), 3.00-2.88 (m, 1 H),2.49 (ddd, J=14.4, 7.4, 5.3 Hz, 1 H), 2.45 (s, 3 H), 2.13-2.00 (m, 1 H),1.11-0.97 (m, 9 H); MS: (ES) m/z calculated for C₂₁H₂₃F₄N₅O [M+H]⁺438.2, found 438.0.

Example 7 Synthesis ofN-[1-(4-fluorophenyl)-5-isopropylpyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

To a solution of 1-(4-fluorophenyl)-5-isopropylpyrazol-4-amine (0.03 g,0.14 mmol) in DMF (1.0 mL) was added2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanoic acid (0.031 g,0.14 mmol), followed by diisopropylethylamine (0.042 g, 0.32 mmol) andHATU (0.057 g, 0.15 mmol). After stirring at room temperature for 1 h,the reaction was diluted with water and the aqueous layer was extractedwith ethyl acetate (2×5 mL). The combined organic layers were washedwith brine, dried (Na₂SO₄), filtered, and concentrated in vacuo. Theresulting crude product was purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to give the title compound asa white solid (0.0145 g, 0.034 mmol, 25%). ¹H NMR (400 MHz, CDCl₃) 7.92(d, J=0.6 Hz, 1 H), 7.46 (q, J=1.2 Hz, 1 H), 7.33-7.24 (m, 2 H),7.21-7.11 (m, 2 H), 6.63 (s, 1 H), 4.92 (q, J=7.3 Hz, 1 H), 2.93 (m, 1H), 2.49 (s, 3 H), 1.89 (d, J=7.3 Hz, 3 H), 1.04 (dd, J=10.3, 7.2 Hz, 6H); MS: (ES) m/z calculated for C₂₀H₂₁F₄N₅O [M+H]⁺ 424.2, found 424.0.

Example 8 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-N-[1-(4-fluorophenyl)-5-isopropylpyrazol-4-yl]butanamide

To a solution of 1-(4-fluorophenyl)-5-isopropylpyrazol-4-amine (0.03 g,0.14 mmol) in DMF (1.0 mL) was added2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]butanoic acid(0.037 g, 0.14 mmol), followed by diisopropylethylamine (0.042 g, 0.32mmol) and HATU (0.057 g, 0.15 mmol). After stirring at room temperaturefor 1 h, the reaction was diluted with water and the aqueous layer wasextracted with ethyl acetate (2×5 mL). The combined organic layers werewashed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo.The resulting crude product was purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to give the titlecompound as a white solid (0.020 g, 0.042 mmol, 31%). ¹H NMR (400 MHz,CDCl₃) δ 8.47 (s, 1 H), 8.04 (s, 1 H), 7.35-7.24 (m, 2 H), 7.21-7.11 (m,2 H), 4.80 (dd, J=8.7, 6.5 Hz, 1 H), 2.98 (m, 1 H), 2.42-2.29 (m, 5 H),1.26 (d, J=7.2 Hz, 3 H), 1.10 (d, J=7.1 Hz, 3 H), 0.92 (t, J=7.3 Hz, 3H); MS: (ES) m/z calculated for C₂₁H₂₂ClF₄N₅O [M+H]⁺ 472.1, found 471.9.

Example 9 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-N-[1-(4-fluorophenyl)-5-isopropylpyrazol-4-yl]propanamide

To a solution of 1-(4-fluorophenyl)-5-isopropylpyrazol-4-amine (0.03 g,0.14 mmol) in DMF (1.0 mL) was added2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]propanoic acid(0.035 g, 0.14 mmol), followed by diisopropylethylamine (0.042 g, 0.32mmol) and HATU (0.057 g, 0.15 mmol). After stirring at room temperaturefor 1 h, the reaction was diluted with water and the aqueous layer wasextracted with ethyl acetate (2×5 mL). The combined organic layers werewashed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo.The resulting crude product was purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to give the titlecompound as a white solid (0.020 g, 0.044 mmol, 32%). ¹H NMR (400 MHz,CDCl₃) δ 8.18 (s, 1 H), 8.02 (s, 1 H), 7.34-7.24 (m, 2 H), 7.21-7.11 (m,2 H), 5.05 (q, J=7.2 Hz, 1 H), 3.03-2.91 (m, 1 H), 2.38 (s, 3 H), 1.90(d, J=7.2 Hz, 3 H), 1.25 (dd, J=13.9, 7.1 Hz, 3 H), 1.08 (d, J=7.2 Hz, 3H); MS: (ES) min calculated for C₂₀H₂₀ClF₄N₅O [M+H]⁺ 458.1, found 457.9.

Example 10 Synthesis ofN-[5-tert-butyl-1-(4-fluorophenyl)pyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

To a solution of 5-tert-butyl-1-(4-fluorophenyl)pyrazol-4-amine (0.03 g,0.13 mmol) in DMF (1.0 mL) was added2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanoic acid (0.029 g,0.13 mmol), followed by diisopropylethylamine (0.033 g, 0.26 mmol) andHATU (0.054 g, 0.14 mmol). After stirring at room temperature for 1 h,the reaction was diluted with water and the aqueous layer was extractedwith ethyl acetate (2×5 mL). The combined organic layers were washedwith brine, dried (Na₂SO₄), filtered, and concentrated in vacuo. Theresulting crude product was purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to give the title compound asa white solid (0.0145 g, 0.034 mmol, 25%). ¹H NMR (400 MHz, CDCl₃) δ7.77 (s, 1 H), 7.45 (q, J=1.2 Hz, 1 H), 7.33-7.24 (m, 2 H), 7.16-7.07(m, 2 H), 6.89 (s, 1 H), 4.90 (d, J=7.3 Hz, 1 H), 2.47 (s, 3 H), 1.86(d, J=7.3 Hz, 3 H), 1.11-1.03 (m, 9 H); MS: (ES) m/z calculated forC₂₁H₂₃F₄N₅O [M+H]⁺ 438.2, found 438.0.

Example 11 Synthesis ofN-[1-(4-chlorophenyl)-5-isopropylpyrazol-4-yl]-2-[5-methyl-3-(trifluoromethyl)-1,2,4-triazol-1-yl]propanamide

To a solution of 1-(4-chlorophenyl)-5-isopropylpyrazol-4-amine (0.054 g,0.23 mmol) in DMF (1.0 mL) was added2-[5-methyl-3-(trifluoromethyl)-1,2,4-triazol-1-yl]propanoic acid (0.051g, 0.23 mmol), followed by diisopropylethylamine (0.059 g, 0.46 mmol)and HATU (0.105 g, 0.28 mmol). After stirring at room temperature for 1h, the reaction was diluted with water and the aqueous layer wasextracted with ethyl acetate (2 k 5 mL). The combined organic layerswere washed with brine, dried (Na₂SO₄), filtered, and concentrated invacuo. The resulting crude product was purified by reverse phase HPLC(C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) to give the titlecompound as a white solid (0.025 g, 0.057 mmol, 25%). ¹H NMR (400 MHz,CDCl₃) δ 8.25 (s, 1 H), 8.03 (d, J=0.6 Hz, 1 H), 7.49-7.41 (m, 2 H),7.32-7.23 (m, 2 H), 5.12 (q, J=7.2 Hz, 1 H), 3.02 (m, 1 H), 2.61 (s, 3H), 1.92 (d, J=7.2 Hz, 3 H), 1.27 (d, J=7.2 Hz, 3 H), 1.10 (d. J=7.2 Hz,3 H); MS: (ES) m/z calculated for C₁₉H₂₀ClF₃N₆O [M+H]⁺ 441.1, found440.9.

Example 12 Synthesis ofN-[1-(4-fluorophenyl)-5-isopropylpyrazol-4-yl]-2-[5-methyl-3-(trifluoromethyl)-1,2,4-triazol-1-yl]propanamide

To a solution of 1-(4-fluorophenyl)-5-isopropylpyrazol-4-amine (0.050 g,0.23 mmol) in DMF (1.0 mL) was added2-[5-methyl-3-(trifluoromethyl)-1,2,4-triazol-1-yl]propanoic acid (0.051g, 0.23 mmol), followed by diisopropylethylamine (0.059 g, 0.46 mmol)and HATU (0.105 g, 0.28 mmol). After stirring at room temperature for 1h, the reaction was diluted with water and the aqueous layer wasextracted with ethyl acetate (2×5 mL). The combined organic layers werewashed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo.The resulting crude product was purified by reverse phase HPLC (C18column, acetonitrile-H₂O with 0.1% TFA as eluent) to give the titlecompound as a white solid (0.03 g, 0.0706 mmol, 31%). ¹H NMR (400 MHz,CDCl₃) δ 8.24 (s, 1 H), 8.04-7.99 (m, 1 H), 7.35-7.25 (m, 2 H),7.22-7.11 (m, 2 H), 5.13 (q, J=7.2 Hz, 1 H), 3.05-2.93 (m, 1 H), 2.62(s, 3 H), 1.93 (d, J=7.2 Hz, 3 H), 1.27 (dd, J=7.2, 2.9 Hz, 3 H), 1.10(d, J=7.1 Hz, 3 H); MS: (ES) m/z calculated for C₁₉H₂₀F₄N₆O [M+H]⁺425.2, found 425.0.

Example 13 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-N-[1-(4-fluorophenyl)-5-isopropylpyrazol-4-yl]-2-methyl-propanamide

To a solution of 1-(4-fluorophenyl)-5-isopropylpyrazol-4-amine (0.03 g,0.14 mmol) in DMF (1.0 mL) was added2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-2-methyl-propanoicacid (0.037 g, 0.14 mmol), followed by diisopropylethylamine (0.042 g,0.32 mmol) and HATU (0.057 g, 0.15 mmol). After stirring at roomtemperature for 1 h, the reaction was diluted with water and the aqueouslayer was extracted with ethyl acetate (2×5 mL). The combined organiclayers were washed with brine, dried (Na₂SO₄), filtered, andconcentrated in vacuo. The resulting crude product was purified byreverse phase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA aseluent) to give the title compound as a white solid (0.025 g, 0.053mmol, 39%). ¹H NMR (400 MHz, CDCl₃) δ 7.91 (d, I=0.6 Hz, 1 H), 7.33-7.25(m, 2 H), 7.20-7.10 (m, 2 H), 6.61 (s, 1 H), 2.92 (m, 1 H), 2.37 (s, 3H), 1.97 (s, 6 H), 1.03 (d, J=7.1 Hz, 6 H); MS: (ES) m/z calculated forC₂₁H₂₂ClF₄N₅O [M+H]⁺ 472.1, found 471.9.

Example 14 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl]-N-[1-(4-fluorophenyl)-5-methyl-1H-imidazol-4-yl]acetamide

a) Acetic anhydride (1.65 mL) was added to 5-methyl-4-nitro-1H-imidazole(254 mg, 2.00 mmol) at room temperature. Nitric acid (70%, 200 μL) wasadded and the reaction was stirred for three days. The reaction was thenpoured onto ice. The product was extracted with dichloromethane threetimes. The combined organic layers was washed with saturated sodiumbicarbonate solution and dried over anhydrous sodium sulfate. Afterremoval of the solvent under reduced pressure, the crude5-methyl-1,4-dinitro-1H-imidazole (235 mg) was dissolved in methanol (3mL) at ambient temperature. A methanol (0.8 mL) solution of4-fluoroaniline (157 μL, 1.63 mmol) was added dropwise and the reactionwas stirred at this temperature for five days. Water was added to thereaction mixture and the product was extracted with dichloromethanethree times. The combined organic layers were dried over anhydroussodium sulfate. After removal of solvent under reduced pressure, thecrude material was purified using silica gel column chromatography(33-60% ethyl acetate in hexanes) to give5-methyl-4-nitro-1-(4-fluorophenyl)-1H-imidazole (157 mg, 0.712 mmol,36% yield over two steps).

b) To a dichloromethane (2 mL) suspension of4-chloro-5-methyl-3-trifluoromethyl-1H-pyrazole-1-acetic acid (186 mg,0.768 mmol), was added oxalyl chloride (134 μL, 1.54 mmol) at ambienttemperature. A catalytic amount of dimethylformamide (1.5 μL) was addedand the reaction was stirred for two hours. Removal of solvent underreduced pressure gave the crude4-chloro-5-methyl-3-trifluoromethyl-1H-pyrazole-1-acetyl chloride whichwas used without any further purification in the next step.

c) Both 5-methyl-4-nitro-1-(4-fluorophenyl)-1H-imidazole (77.8 mg, 0.352mmol) and 4-chloro-5-methyl-3-trifluoromethyl-1H-pyrazole-1-acetylchloride (crude, approximately 0.768 mmol) were dissolved in a mixtureof ethyl acetate (5 mL) and tetrahydrofuran (5 mL). After flushing thereaction mixture with nitrogen, palladium on carbon (10%, wet, 23.3 mg)was added. The reaction mixture was hydrogenated using a Parr Apparatusfor one and half hours at 40 psi. The reaction mixture was then filteredto remove palladium on carbon and after removal of the solvents underreduced pressure, the crude material was purified using silica gelcolumn chromatography (1.3-2.6% methanol in ethyl acetate with 0.05%aqueous ammonia) to afford2-[4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl]-N-[1-(4-fluorophenyl)-5-methyl-1H-imidazol-4-yl]acetamide(13.6 mg, 0.0327 mmol, 9% yield). ¹H NMR (400 MHz, CDCl₃) δ 7.73 (br, 1H), 7.39 (s, 1 H), 7.24-7.29 (m, 2 H), 7.19 (dd, J=8.4, 8.4 Hz, 1 H),4.94 (s, 2 H), 2.35 (s, 3 H), 2.04 (s, 3 H); MS: (ES) m % z calculatedfor C₁₇H₁₄N₅OClF₄ [M+H]⁺ 416.1, found 416.1.

Example 15N-[1-(4-Chlorophenyl)-5-isopropyl-pyrazol-4-yl]-3-methyl-1-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyridine-5-carboxamide

a) To 2-methyl-4-(trifluoromethyl)-1H-imidazole (1 g, 6.7 mmol) wasadded CHCl₃ (4.5 mL) and AcOH (13.5 mL) followed by I₂ (1.78 g, 6.99mmol) and HIO₄.2H₂O (1.52 g, 6.7 mmol), and the resulting reactionmixture was stirred at 60° C. for 3 h. The reaction mixture was thenpoured into ice cold 10% aqueous sodium bisulphite solution (100 mL) andthe aqueous layer was extracted with EtOAc (3:100 mL). The combinedorganic layers were washed with brine (100 mL), dried (MgSO₄) andconcentrated in vacuo to obtain5-iodo-2-methyl-4-(trifluoromethyl)-1H-imidazole (1.98 g) which was usedas such in next step without further purification.

b) To a solution of 5-iodo-2-methyl-4-(trifluoromethyl)-1H-imidazole(1.98 g, 7.2 mmol) in DME (20 mL) and H₂O (4 mL) was added K₂CO₃ (10 g,71.9 mmol), trivinylboronic anhydride pyridine complex (1.73 g, 7.19mmol) and Pd(PPh₃)₄ (830 mg, 0.719 mmol). The resulting reaction mixturewas stirred at 80° C. for 3 h. MeOH (100 mL) was then added to thereaction mixture and the mixture was filtered, concentrated in vacuo,and purified by flash chromatography (SiO₂, 10% MeOH/CH₂Cl₂) to obtain2-methyl-4-(trifluoromethyl)-5-vinyl-1H-imidazole (911 mg, 5.17 mmol,72% yield) as a brown syrup.

c) To a solution of 2-methyl-4-(trifluoromethyl)-5-vinyl-1H-imidazole(911 mg, 5.17 mmol) was added DMF (7 mL) and THF (3 mL), followed byK₂CO₃ (1.43 g, 10.35 mmol) and ethyl bromo acetate (687 μL, 6.2 mmol).The resulting mixture was then stirred overnight at room temperature.Water (50 mL) was added and the mixture was extracted with EtOAc (3×100mL). The combined EtOAc layers were dried (MgSO₄), concentrated invacuo, and purified by flash chromatography (SiO₂, 60% EtOAc/hexanes) toobtain the desired ethyl2-[2-methyl-4-(trifluoromethyl)-5-vinyl-imidazol-1-yl]acetate (796 mg,3.03 mmol, 59% yield) as a yellow oil.

d) To a cooled (−78° C.) solution of ethyl2-[2-methyl-4-(trifluoromethyl)-5-vinyl-imidazol-1-yl]acetate (790 mg,3.01 mmol) in THF (6 mL) under nitrogen atmosphere was added LDA (2 M, 3mL, 6.03 mmol) drop wise. After stirring the reaction mixture at −78° C.for 15 min, allyl bromide (521 μL, 6.03 mmol) was added slowly and thereaction mixture was allowed to warm to room temperature and stirred for2 h. A saturated NH₄Cl solution (20 mL) was added to the reactionmixture at 0° C. and the mixture was then extracted with EtOAc (2×100mL). The combined EtOAc layers were then washed with brine (50 mL),dried (MgSO₄), concentrated in vacuo, and purified by flashchromatography (SiO₂, 80% EtOAc/hexanes) to obtain ethyl2-[2-methyl-4-(trifluoromethyl)-5-vinyl-imidazol-1-yl]pent-4-enoate (127mg, 0.42 mmol, 14% yield).

e) To ethyl2-[2-methyl-4-(trifluoromethyl)-5-vinyl-imidazol-1-yl]pent-4-enoate (127mg, 0.42 mmol) was added EtOH (2 mL) and 5 N NaOH (0.5 mL), and theresulting solution was stirred at room temperature for 1 h. 12 N HCl wasadded slowly at room temperature until the solution reached pH 2,followed by concentration in vacuo to obtain2-[2-methyl-4-(trifluoromethyl)-5-vinyl-imidazol-1-yl]pent-4-enoic acid(100 mg, crude) which was used as such in the next step without furtherpurification.

f) To 2-[2-methyl-4-(trifluoromethyl)-5-vinylimidazol-1-yl]pent-4-enoicacid (120 mg, 0.42 mmol) was added DMF (3 mL), Et₃N (500 μL, excess),1-(4-chlorophenyl)-5-isopropylpyrazol-4-amine (100 mg, 0.42 mmol), andHATU (250 mg, excess) and the resulting mixture was stirred at roomtemperature for 1 h. Water (20 mL) was added and the mixture wasextracted with EtOAc (2×50 mL). The combined organics were then dried(MgSO₄), concentrated in vacuo, and purified by flash chromatography(SiO₂, 80% EtOAc/hexanes) to obtainN-[1-(4-chlorophenyl)-5-isopropylpyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)-5-vinylimidazol-1-yl]pent-4-enamide(147 mg, 0.3 mmol, 70% yield).

g) ToN-[1-(4-chlorophenyl)-5-isopropylpyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)-5-vinyl-imidazol-1-yl]pent-4-enamide(147 mg, 0.298 mmol) was added CH₂Cl₂ (10 mL) andbenzylidene-bis(tricyclohexylphosphine)dichlororuthenium (Grubb's firstgeneration catalyst, 122.6 mg, 0.149 mmol), and the resulting mixturewas stirred at 45° C. for 3 h. The reaction mixture was then directlyadsorbed on SiO₂ and purified by flash chromatography (SiO₂, 80%EtOAc/hexanes) to furnishN-[1-(4-chlorophenyl)-5-isopropylpyrazol-4-yl]-3-methyl-1-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyridine-5-carboxamide(30 mg, 0.064 mmol, 22% yield).

h) ToN-[1-(4-chlorophenyl)-5-isopropylpyrazol-4-yl]-3-methyl-1-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyridine-5-carboxamide(30 mg, 0.064 mmol) was added EtOH (10 mL), 12 N HCl (4 drops), and PtO₂(30 mg). The resulting suspension was evacuated with hydrogen gas twiceand stirred under hydrogen gas (55 psi) on Parr shaker for 25 min. thereaction mixture was then filtered through a syringe filter,concentrated in vacuo, and purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to giveN-[1-(4-chlorophenyl)-5-isopropyl-pyrazol-4-yl]-3-methyl-1-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyridine-5-carboxamide(10 mg, 0.0172 mmol, 27% yield) as a TFA salt. ¹H NMR (400 MHz,Methanol-d₄) δ 7.62 (s, 1 H), 7.57 (d, J=11.76 Hz, 2 H), 7.42 (d,J=11.76 Hz, 2 H), 5.20-5.22 (m, 1 H), 3.0-3.18 (m, 2 H), 2.8-2.95 (m, 1H), 2.40 (s, 3 H), 2.32-2.55 (m, 2 H), 1.80-2.05 (m, 2 H), 1.27 (d,J=23.4 Hz, 3 H), 1.24 (d, J=23.4 Hz, 3 H); MS: (ES) m/z calculated forC₂₂H₂₃ClF₃N₅O [M+H]+ 466.9, found 466.1.

Example 15 Synthesis of2-(4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)-N-(1-(4-fluorophenyl)-5-isopropyl-1H-pyrazol-4-yl)-6-hydroxyhexanamide

a) Trimethylaluminum (1.5 mL, 3 mmol, 2 M solution in toluene) was addedportionwise to a solution of1-(4-fluorophenyl)-5-isopropyl-1H-pyrazol-4-amine (329 mg, 1.5 mmol) andα-bromocaprolactone (319 mg, 1.65 mmol) in anhydrous dichloroethane (7.5mL) under nitrogen at room temperature. After stirring for 1 h, thereaction was diluted with saturated NH₄Cl and the mixture was furtherdiluted with 20 mL of EtOAc and 1.5 mL of 6 N HCl. The organic layer waswashed with water and brine, dried over MgSO₄, filtered, andconcentrated in vacuo. The crude material was used without furtherpurification.

b) The residue from step a (310 mg, 0.75 mmol) and4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazole (277 mg, 1.5 mmol) wasdissolved in DMF (1.5 mL) and treated with K₂CO₃ (311 mg, 2.25 mmol).After stirring at 60° C. for 2.5 h, the reaction mixture was dilutedwith EtOAc (20 mL) and washed with water and brine. The organic layerwas dried on MgSO₄, filtered, and concentrated in vacuo to give aresidue that was purified by flash chromatography (SiO₂, 35-100%EtOAc/hexanes) to give the titled compound (300 mg, 0.58 mmol, 78%) as acolorless oil that solidified on cooling. ¹H NMR (400 MHz, CDCl₃) δ 8.37(s, 1 H), 8.03 (s, 1 H), 7.32-7.27 (m, 2 H), 7.18-7.13 (m, 2 H), 4.89(dd, J=9.4, 6.2 Hz, 1 H), 3.65 (ddd, J=12.6, 10.6, 6.7 Hz, 2 H),3.00-2.93 (m, 1 H), 2.37 (s, 3 H), 1.65-1.56 (m, 4 H), 1.48-1.37 (m, 1H), 1.32-1.28 (m, 1 H), 1.25 (d, J=7.0 Hz, 3 H), 1.08 (d, J=7.0 Hz, 3H); MS: (ES) m/z calculated for C₂₃H₂₇ClF₄N₅O₂[M+H]⁺ 516.2, found 516.1.

Example 15 Synthesis of(S)—N-(1-(4-chlorophenyl)-5-isopropyl-1H-pyrazol-4-yl)-2-(2-methyl-4-(trifluoromethyl)-1H-imidazol-1-yl)propanamide

a) A solution of ethyl 2-bromopropionate (3.98 g, 22 mmol),4-chloro-5-methyl-3-(trifluoromethyl)-1H-pyrazole (3.00 g, 20 mmol), andK₂CO₃ (5.53 g, 40 mmol) in THF/DMF (2:1, 39 mL) was allowed to stir at45° C. for 16 h. The mixture was then concentrated in vacuo and dilutedwith EtOAc (70 mL). The organic layer was washed with water and brine,dried over MgSO₄, filtered, and concentrated in vacuo to give theproduct (5.3 g) as a colorless oil that was used without furtherpurification.

b) The crude material from step a was dissolved in THF (40 mL) andtreated with 2 M LiOH (15 mL, 30 mmol). The slurry was heated to 60° C.for 1 h and then concentrated. The residue was diluted with water (20mL) and adjusted to pH 2 with H₂SO₄ and NaOH, providing the desired acidas a colorless solid (3.03 g, 13.6 mmol, 68% over two steps).

c) The acid intermediate (1.00 g, 4.5 mmol) from step b and(S)-phenylglycinol (679 mg, 4.95 mmol) were slurried in THF (22 mL) andEt₃N (1.25 mL, 9 mmol). HATU (1.88 g, 4.95 mmol) was added and theslurry was stirred for 4 h. The volatiles were removed in vacuo and theresidue diluted in EtOAc. The organic layer was washed with 3 M KOH(2×10 mL) and brine and was then loaded onto silica gel. The crudematerial was purified by flash chromatography (SiO₂, 3-4%methanol/CH₂Cl₂) to give two diastereomeric products as colorlesssolids. The first eluting isomer (500 mg) was obtained in >99:1diastereomeric ratio (by ¹H NMR).

d) The first eluting product from step c (484 mg, 1.4 mmol) wasdissolved in dioxane (5.6 mL) and treated with 6 M H₂SO₄ (3.5 mL, 21mmol). The slurry was heated at 80° C. for 6 h and then cooled. Thecrude residue was then purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent). The resulting lactone.TFAsalt was neutralized and salted with HCl and dried to provide theenantiomerically enriched acid as a colorless solid (301 mg, 1.16 mmol,83%).

e) To a solution of the acid intermediate from step d (23.6 mg, 0.1mmol) and 1-(4-chlorophenyl)-5-isopropyl-1H-pyrazol-4-amine (16.6 mg,0.064 mmol) in DMF (0.5 mL) was added Et₃N (20 μL, 0.13 mmol) and HATU(31.6 mg, 0.083 mmol). After stirring 35 min, the slurry wasconcentrated under reduced pressure and the residue was purified byreverse phase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA aseluent). The residue was neutralized and salted with HCL to provide thetitled compound hydrochloride salt as a colorless solid (12.3 mg). ¹HNMR (400 MHz, CD₃OD) δ 8.28 (d, J=1.6 Hz, 1 H), 7.63 (s, 1 H), 7.57(ddd, J=9.7, 5.1, 3.1 Hz, 2 H), 7.41 (ddd, J=9.7, 5.0, 3.1 Hz, 2 H),5.38 (q, J=7.0 Hz, 1 H), 3.03 (hept, J=7.0 Hz, 1 H), 2.65 (s, 3 H), 1.91(d, J=7.0 Hz, 3 H), 1.23 (d, J=7.1 Hz, 3 H), 1.22 (d, J=7.0 Hz, 3 H);MS: (ES) m/z calculated for C₂₀H₂₂ClF₃N₅O [M+H]⁺ 440.1, found 439.9.Retention time on chiral HPLC: 5.9 min (RegisPack cat#783104, 25 cm×4.6mm, 5 micron; eluent: 0.1% diethylamine/IPA, 1.0 ml/min). The er isdetermined to be 20:1 with the (R)-enantiomer having a retentime of 3.4min. Absolute configuration of the (R)-enantiomer was confirmed by anindependent synthesis from methyl (L)-lactate.

Example 16 Synthesis of(S)—N-(1-(4-fluorophenyl)-5-isopropyl-1H-pyrazol-4-yl)-2-(2-methyl-4-(trifluoromethyl)-1H-imidazol-1-yl)propanamidetrifluoracetic acid salt

The titled compound was prepared using the procedure as described inExample 15, substituting1-(4-chlorophenyl)-5-isopropyl-1H-pyrazol-4-amine for1-(4-fluorophenyl)-5-isopropyl-1H-pyrazol-4-amine in step e. The productwas isolated as the TFA salt ¹H NMR (400 MHz, CD₃OD) δ 7.85 (d, J=1.2Hz, 1 H), 7.59 (d, J=1.2 Hz, 1 H), 7.47-7.41 (m, 2 H), 7.32-7.26 (m, 2H), 5.21 (q, J=7.0 Hz, 1 H), 2.99 (hept, J=7.0 Hz, 1 H), 2.50 (s, 3 H),1.83 (d, J=7.1 Hz, 3 H), 1.19 (d, J=6.6 Hz, 3 H), 1.18 (d, J=6.7 Hz, 3H); MS: (ES) m/z calculated for C₂₀H₂₂F₄N₅O [M+H]⁺ 424.2, found 423.9.Retention time on chiral HPLC: 13.1 min (RegisPack cat#783104, 25 cm×4.6mm, 5 micron; eluent: 0.1% diethylamine/IPA, 0.4 ml/min). The er isdetermined to be 40:1 with the (R)-enantiomer having a retention time of8.2 min.

Example 17 Synthesis ofN-[5-ethoxy-1-(4-fluorophenyl)pyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

a) A mixture of 4-iodofluorobenzene (2.42 g, 11 mmol),4-nitro-1H-pyrazole (1.00 g, 10 mmol), 8-hyroxyquinoline (0.15 g, 1mmol), CuI (0.192 g, 1 mmol), and potassium carbonate (2.78 g, 20 mmol)in DMSO (20 mL) was heated at 135° C. overnight. After cooling to roomtemperature, the reaction mixture was diluted with 30 mL of water andextracted with ethyl acetate. The organic layer was washed with aqueoussaturated sodium bicarbonate, dried (Na₂SO₄), filtered, and concentratedin vacuo. Purification by flash chromatography (SiO₂, 10%-20%. EtOAc inhexanes) gave 1.02 g (4.9 mmol, 49%) of the desired product.

b) To a stirred solution of 1-(4-fluorophenyl)-4-nitropyrazole (1.02 g,4.9 mmol) in 10 mL of THF was added LiHMDS (1 M in THF, 5.8 mL, 5.8mmol) slowly at −78° C. under nitrogen. After stirring 30 minutes,1,1,1,2,2,2-hexachloroethane (1.31 g, 5.5 mmol) in 6 mL of THF was addeddropwise. The reaction mixture was stirred for 1 h followed by quenchingwith 20 mL of aqueous saturated NH₄Cl. The reaction mixture was thenwarmed to room temperature and extracted with EtOAc. The organic layerwas dried (Na₂SO₄), filtered, and concentrated in vacuo. Purification byflash chromatography (SiO₂, 5%-15% EtOAc in hexanes) provided 0.613 g ofthe desired product (2.5 mmol, 52%).

c) A mixture of 5-chloro-1-(4-fluorophenyl)-4-nitropyrazole (0.121 g,0.5 mmol) and sodium ethoxide (0.137 g, 2 mmol) in 2 mL of EtOH washeated at 75° C. overnight. After cooling to room temperature, thereaction mixture was diluted with 20 mL of aqueous saturated sodiumbicarbonate and extracted with EtOAc. The organic layer was washed withbrine, dried (Na₂SO₄), filtered, and concentrated in vacuo. The crudematerial was used directly in the next step.

d) A mixture of the crude product from step c, iron (0.114 g, 2 mmol),and 100 μL of aqueous 6 N HCl in 2 mL of EtOH was heated at 80° C. for20 minutes. After cooling to room temperature, the reaction mixture wasdiluted with 20 mL of aqueous saturated sodium bicarbonate and 40 mL ofEtOAc. The resulting suspension was stirred for 10 minutes then filteredthrough a pad of Celite. The organic layer was separated, washed withbrine, dried (Na₂SO₄), filtered, and concentrated in vacuo. The crudematerial was used directly in the next step.

e) A mixture of the crude product from step d,2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanoic acid (0.112 g,0.5 mmol), HATU (0.191 g, 0.5 mmol), and 100 μL of Et₃N in 1 mL ofCH₂Cl₂ was stirred at room temperature. After 30 minutes, the reactionmixture was diluted with 10 mL of aqueous saturated sodium bicarbonateand extracted with EtOAc. The organic layer was washed with brine, dried(Na₂SO₄), filtered, and concentrated in vacuo. Purification by reversephase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA as eluent)followed by flash chromatography (SiO₂, 60%-100% EtOAc in hexanes)afforded the title compound as a colorless solid (0.020 g, 0.047 mmol,2.4% for 5 steps). ¹H NMR (400 MHz, CDCl₃) 7.85 (s, 1 H), 7.67-7.55 (m,2 H), 7.44 (s, 1 H), 7.31-7.08 (m, 2 H), 6.82 (s, 1 H), 4.88 (q, J=7.2Hz, 1 H), 3.84 (q, J=7.1 Hz, 2 H), 2.45 (s, 3 H), 1.85 (d, J=7.2 Hz, 3H), 1.65 (s, 3 H), 1.17 (t, J=7.1 Hz, 3 H). MS: (ES) m/z calculated forC₁₉H₁₉F₄N₅O₂ [M+H]⁺ 426.2, found 426.1.

Example 18 Synthesis ofN-[5-chloro-1-(4-fluorophenyl)pyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

A sample of 5-chloro-1-(4-fluorophenyl)-4-nitropyrazole generated in theprevious example was subsequently carried through steps d and e. Thetitled compound was isolated during the purification described inExample 17, providing 2.5 mg as a colorless solid. ¹H NMR (400 MHz,CDCl₃) δ 8.13 (s, 1 H), 7.48-7.52 (m, 2 H), 7.45 (s, 1 H), 7.16-7.21 (m,2 H), 6.72 (s, 1 H), 4.92 (q, J=7.2 Hz, 1 H), 2.49 (s, 3 H), 1.88 (d,J=7.2 Hz, 3 H). MS: (ES) m/z calculated for C₁₇H₁₄ClF₄N₅O [M+H]⁺ 416.1,found 416.1.

Example 19 Synthesis ofN-[5-isopropoxy-1-(4-fluorophenyl)pyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

a) To a stirred solution of isopropanol (0.12 g, 2 mmol) in 1 mL of NMPwas added NaH (0.085 g, 2 mmol) portionwise at 0° C. The reactionmixture was warmed to room temperature for 10 minutes before theaddition of 5-chloro-1-(4-fluorophenyl)-4-nitropyrazole (0.24 g, 1 mmol)in one portion. The reaction slurry was then heated at 100° C. for 3 h.After cooling to room temperature, the reaction was quenched withaqueous saturated sodium bicarbonate and extracted with EtOAc. Theorganic layer was dried (Na₂SO₄), filtered, and concentrated in vacuo.The crude material was used directly in the next step.

b) A mixture of the crude product from step a, iron (0.23 g, 4 mmol),and 100 μL of aqueous 6 N HCl in 2 mL of EtOH was heated at 80° C. for20 minutes. After cooling to room temperature, the reaction mixture wasdiluted with 20 mL of aqueous saturated sodium bicarbonate and 40 mL ofEtOAc. The resulting suspension was stirred for 10 minutes then filteredthrough a pad of Celite. The organic layer was separated, washed withbrine, dried (Na₂SO₄), filtered, and concentrated in vacuo. The crudematerial was used directly in the next step.

c) A mixture of crude 1-(4-fluorophenyl)-5-isopropoxy-pyrazol-4-amine(prepared in step b, 0.032 g, 0.13 mmol),2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanoic acid (0.031 g,0.13 mmol), HATU (0.057 g, 0.13 mmol), and 100 μL of Et₃N in 1 mL ofCH₂Cl₂ was stirred at room temperature. After 30 min, the reactionmixture was diluted with 10 mL of aqueous saturated sodium bicarbonateand extracted with EtOAc. The organic layer was washed with brine, dried(Na₂SO₄), filtered, and concentrated in vacuo. Purification by reversephase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA as eluent)provided the title compound as a colorless solid (0.030 g, 0.068 mmol,520% for 3 steps). ¹H NMR (400 MHz, CD₃OD) δ 7.81 (s, 1 H), 7.70 (s, 1H), 7.68-7.57 (m, 2 H), 7.30-7.19 (m, 2 H), 5.16 (q, J=6.5 Hz, 1 H),4.26 (he, J=6.1 Hz, 1 H), 2.47 (s, 3 H), 1.80 (d, J=6.5 Hz, 3 H), 1.14(d, J=6.1, 6 H). MS: (ES) m/z calculated for C₂₀H₂₁F₄N₅O₂ [M+H]⁺ 439.2,found 439.4.

Example 20 Synthesis ofN-[5-(dimethylamino)-1-(4-fluorophenyl)pyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

a) A mixture of 5-chloro-1-(4-fluorophenyl)-4-nitropyrazole (0.20 g, 0.8mmol) and dimethylamine (2 M in water, 0.80 mL, 1.6 mmol) in 1 mL of DMFwas heated at 80° C. After cooling to room temperature, the reaction wasdiluted with 20 mL of water and extracted with EtOAc. The organic layerwas washed with brine, dried (Na₂SO₄), filtered, and concentrated invacuo. The crude material was used directly in the next step.

b) A mixture of crude product from step a, iron (0.14 g, 2.5 mmol), and100 μL of aqueous 6 N HCl in 1 mL of EtOH was heated at 80° C. for 20minutes. Upon cooling to room temperature, the reaction mixture wasdiluted with 20 mL of aqueous saturated sodium bicarbonate, and 40 mL ofEtOAc. The resulting suspension was stirred for 10 minutes and thenfiltered through celite. The organic layer was separated, washed withbrine, dried (Na₂SO₄), filtered, and concentrated in vacuo. The crudematerial was used directly in the next step.

c) A mixture of crude2-(4-fluorophenyl)-N3,N3-dimethyl-pyrazole-3,4-diamine (from step b,0.042 g, 0.18 mmol),2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanoic acid (0.060 g,0.27 mmol), HATU (0.10 g, 0.27 mmol), and 100 μL of Et₃N in 1 mL ofCH₂Cl₂ was stirred at room temperature. After 30 minutes, the reactionmixture was diluted with 10 mL of aqueous saturated sodium bicarbonateand extracted with EtOAc. The organic layer was washed with brine, dried(Na₂SO₄), filtered, and concentrated in vacuo. Purification by reversephase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA as eluent)afforded the title compound as a colorless solid (0.019 g, 0.045 mmol,25% for 3 steps). ¹H NMR (400 MHz, CDCl₃) δ 7.77 (s, 1 H), 7.53-7.43 (m,2 H), 7.27 (s, 1 H), 7.17-7.08 (m, 2 H), 6.91 (s, 1 H), 4.90 (q, J=7.2Hz, 1 H), 2.53 (s, 6 H), 2.45 (s, 3 H), 1.85 (d, J=7.2 Hz, 3 H). MS:(ES) m/z calculated for C₁₉H₂₀F₄N₆O₁ [M+H]⁺ 424.2, found 424.1.

Example 21 Synthesis ofN-[1-(4-chlorophenyl)-5-isopropyltriazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

a) To a cooled (0° C.) slurry of 4-chloroaniline (0.25 g, 2 mmol) in 20mL of aqueous 4 N HCl was added a solution of sodium nitrite (0.14 g, 2mmol) in 200 μL of H₂O. After 10 minutes, sodium azide (0.16 g, 2.4mmol) was added and the reaction mixture was stirred at roomtemperature. After 2 h, the reaction mixture was extracted with ethylacetate. The organic layer was washed with aqueous saturated sodiumbicarbonate, dried (Na₂SO₄), filtered, and concentrated in vacuo. Thecrude material was used directly in the next step.

b) To stirring solution of methyl 4-methyl-3-oxo-pentanoate (341 μL, 2.4mmol) in 2 mL of MeOH was added NaOMe at 0° C. After 5 minutes, theresidue from step a in 1 mL of MeOH was added in one portion. Thereaction mixture was then allowed to stir at room temperature overnight.The mixture was then diluted with 20 mL of aqueous saturated sodiumbicarbonate and extracted with EtOAc. The organic layer was washed withbrine, dried (Na₂SO₄), filtered, and concentrated in vacuo. Purificationby flash chromatography (SiO₂, 10%-20% EtOAc in hexanes) gave 0.11 g ofthe desired product (0.39 mmol, 20%).

c) A mixture of methyl1-(4-chlorophenyl)-5-isopropyltriazole-4-carboxylate (0.11 g, 0.39 mmol)and lithium hydroxide in 4 mL of THF and 1 mL of H₂O was heated at 60°C. After 2 h, the slurry was cooled to room temperature, adjusted to pH5, and extracted with EtOAc. The organic layer was washed with brine,dried (Na₂SO₄), filtered, and concentrated in vacuo. The crude materialwas used directly in the next step.

d) To the product from step c in 1 mL of CH₂Cl₂ at 0° C. was addedoxalyl chloride (67 μL, 0.78 mmol) and two drops of DMF. After 5minutes, the reaction mixture was warmed to room temperature. After 2 h,the reaction slurry was concentrated in vacuo. The crude material wasused directly in the next step.

e) The product from step d was diluted in 2 mL of acetone, cooled to 0°C., and treated with NaN₃ (1 g). The reaction slurry was warmed to roomtemperature for 10 minutes. The slurry was then diluted with 10 mL ofaqueous saturated sodium bicarbonate and extracted with EtOAc. Theorganic layer was washed with brine, dried (Na₂SO₄), filtered, andconcentrated in vacuo. The crude material was used directly in the nextstep.

f) A solution of the product from step e in 3 mL of toluene was heatedat 100° C. for 2 h. To this solution was added aqueous 8 N HCl (200 μL,1.6 mmol) at 100° C. and the mixture was stirred for another 10 minutes.After cooling to room temperature, the reaction mixture was diluted with20 mL of aqueous saturated sodium bicarbonate and extracted with EtOAc.The organic layer was washed with brine, dried (Na₂SO₄), filtered, andconcentrated in vacuo. Purification by flash chromatography (SiO₂,10%-40% EtOAc in hexanes) gave 0.05 g of desired aniline product (0.21mmol, 54%).

g) A mixture of 1-(4-chlorophenyl)-5-isopropyltriazol-4-amine (from stepf, 0.025 g, 0.11 mmol),2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanoic acid (0.031 g,0.13 mmol), HATU (0.057 g, 0.13 mmol), and 100 μL of Et₃N in 1 mL ofCH₂Cl₂ was stirred at room temperature. After 30 minutes, the reactionmixture was diluted with 10 mL of aqueous saturated sodium bicarbonateand extracted with EtOAc. The organic layer was washed with brine, dried(Na₂SO₄), filtered, and concentrated in vacuo. Purification by reversephase HPLC (C18 column, acetonitrile-H₂O with 0.1% TFA as eluent)provided the title compound as a colorless solid (0.0051 g, 0.011 mmol,10%). ¹H NMR (400 MHz, CDCl₃) δ 9.14 (s, 1 H), 7.65 (s, 1 H), 7.54 (d,J=8.3 Hz, 2 H), 7.34 (d, J=8.3 Hz, 2 H), 5.26 (q, J=7.4 Hz, 1 H),3.0-2.98 (m, 1 H), 2.62 (s, 3 H), 1.86 (d, J=7.4 Hz, 3 H), 1.14 (t,J=6.4 Hz, 6 H). MS: (ES) m/z calculated for C₁₉H₂₀ClF₃N₆O [M+H]⁺ 441.1,found 441.2.

Example 22 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-N-[1-(4-fluorophenyl)pyrazol-4-yl]acetamide

a) A mixture of 4-fluorobenzeneboronic acid (2.47 g, 17.7 mmol),4-nitro-1H-pyrazole (1.00 g, 8.84 mmol), copper (II) acetate (2.41 g,13.3 mmol) and pyridine (2.86 mL, 35.4 mmol) in CH₂Cl₂ (40 mL) wasstirred at room temperature overnight. The reaction mixture was filteredand diluted with water (40 mL). The organic layer was separated, driedover anhydrous sodium sulfate, concentrated in vacuo, and purified byflash chromatography (SiO₂, 0-20% EtOAc/hexanes gradient elution) togive 1-(4-fluorophenyl)-4-nitropyrazole as a white solid (0.820 g, 22%).

b) A mixture of 1-(4-fluorophenyl)-4-nitropyrazole (0.820 g, 3.96 mmol)and 10% Pd/C (0.133 g, 50% wet by wt) in EtOH (50 mL) was fitted onto aParr apparatus and agitated under H₂ at 40 psi for 40 min. The reactionmixture was then filtered through filter paper. The filtrate wascollected and concentrated in vacuo to give1-(4-fluorophenyl)pyrazol-4-amine as an oil (0.675 g, 96%).

c) To a mixture of2-[4-chloro-3-methyl-5-(trifluoromethyl)pyrazol-1-yl]acetic acid (0.150g, 0.62 mmol) in CH₂Cl₂ (2 mL) was added oxalyl chloride (0.15 mL, 1.75mmol) and DMF (1 drop). The mixture was stirred for 30 min at roomtemperature and was then concentrated in vacuo. The solid obtained wastransferred to another flask containing1-(4-fluorophenyl)pyrazol-4-amine (0.080 g, 0.45 mmol) and NEt₃ (0.25mL, 1.8 mmol) in CH₂Cl₂ (3 mL). The mixture was stirred for 30 min atroom temperature, treated with water (50 mL), and extracted with ethylacetate (50 mL). The organic layer was separated, dried over anhydroussodium sulfate, concentrated in vacuo, and purified by flashchromatography (SiO₂, 0-50% EtOAc/hexanes gradient elution) to give thetitle compound as a white solid (0.084 g, 47%). ¹H NMR (400 MHz CDCl₃) δ8.40 (s, 1 H), 7.71 (s, 1 H), 7.69 (m, 2 H), 7.19 (dd, J=8.4, 8.4 Hz, 2H), 5.07 (s, 2 H), 2.34 (s, 3 H); MS: (ES) m/z calculated forC₁₆H₁₂ClF₄N₅O [M+H]⁺ 402.0, found 402.0.

Example 23 Synthesis of2-[4-chloro-5-methyl-3-(trifluoromethyl)pyrazol-1-yl]-N-[1-(4-fluorophenyl)pyrazol-4-yl]-N-methyl-acetamide

a) A solution of 1-(4-fluorophenyl)pyrazol-4-amine (0.59 g, 3.3 mmol) informic acid (15 mL) was heated at 80° C. for 2 h. The reaction mixturewas cooled to room temperature, concentrated in vacuo, and purified byflash chromatography (SiO₂, 0-100% EtOAc/CH₂Cl₂ gradient elution) togive N-[1-(4-fluorophenyl)pyrazol-4-yl]formamide (0.490 g, 71%).

b) A mixture of N-[1-(4-fluorophenyl)pyrazol-4-yl]formamide (0.275 g,1.33 mmol) and LiAlH₄ (1.33 mL, 2.66 mmol, 2 M in THF) in THF (5 mL) washeated at 45° C. for 1 h. The reaction mixture was cooled to roomtemperature and diluted with 5 mL of concentrated ammonium hydroxide and80 mL of 20% MeOH/CH₂Cl₂. The organic layer was separated, dried overanhydrous sodium sulfate, filtered, concentrated in vacuo, and purifiedby flash chromatography (SiO₂, 0-100% EtOAc/CH₂Cl₂ gradient elution) togive 1-(4-fluorophenyl)-N-methylpyrazol-4-amine as an oil (0.23 g,900/%).

c) To a solution of2-[4-chloro-3-methyl-5-(trifluoromethyl)pyrazol-1-yl]acetic acid (0.070g, 0.29 mmol) in CH₂Cl₂ (2 mL) was added oxalyl chloride (0.10 mL, 1.16mmol) and DMF (1 drop). The mixture was stirred for 30 min at roomtemperature and concentrated in vacuo. The solid obtained wastransferred to another flask containing1-(4-fluorophenyl)-N-methylpyrazol-4-amine (0.056 g, 0.29 mmol) and NEt₃(0.20 mL, 1.4 mmol) in CH₂Cl₂ (3 mL). The reaction mixture was stirredfor 30 min at room temperature, quenched with water (50 mL), andextracted with ethyl acetate (50 mL). The organic layer was separated,dried over anhydrous sodium sulfate, concentrated in vacuo, and purifiedby flash chromatography (SiO₂, 0-50% EtOAc/CH₂Cl₂ gradient elution) togive the title compound (0.045 g, 37%). ¹H NMR (400 MHz, CDCl₃) δ 8.01(s, 1 H), 7.70 (s, 1 H), 7.64 (m, 2 H), 7.17 (dd, J=8.4, 8.4 Hz, 2 H),4.80 (s, 2 H), 3.28 (s, 3 H); MS: (ES) m, calculated for C₁₇H₁₄ClF₄N₅O[M+H]⁺ 416.1, found 416.1.

Example 24 Synthesis ofN-[1-(4-chlorophenyl)-5-isopropylpyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

a) A mixture of methyl 4-methyl-3-oxovalerate (15.0 g, 104 mmol) andN,N-dimethylfomamide dimethyl acetal (69 g, 580 mmol) was heated at 100°C. for 1 h. The reaction mixture was cooled to room temperature,concentrated in vacuo, and azeotroped with toluene. The obtained oilyresidue was used without further purification.

b) A mixture of the intermediate from step a (approx. 104 mmol),4-chlorophenylhydrazine hydrochloride (18.6 g 104 mmol), and K₂CO₃ (28.8g, 208 mmol) in DMF (150 mL) was heated at 100° C. for 1 h. The reactionmixture was cooled to room temperature, diluted with saturated aqueousNH₄Cl (400 mL), and extracted with EtOAc (600 mL). The organic layer wasseparated, dried over anhydrous sodium sulfate, concentrated in vacuo,and purified by flash chromatography (SiO₂, 0-10% EtOAc/CH₂Cl₂ gradientelution) to give methyl1-(4-chlorophenyl)-5-isopropyl-pyrazole-4-carboxylate (26.0 g, 90%).

c) A mixture of methyl1-(4-chlorophenyl)-5-isopropyl-pyrazole-4-carboxylate (26.0 g, 96.7mmol) and lithium hydroxide monohydrate (10.0 g, 238 mml) in MeOH (60mL), THF (60 mL), and H₂O (30 mL) was heated at 80° C. for 3 h. Thereaction mixture was cooled to room temperature, acidified with 1 Naqueous HCl, and extracted with EtOAc (600 ml). The organic layer wasseparated, dried over anhydrous sodium sulfate, and concentrated invacuo to yield 1-(4-chlorophenyl)-5-isopropylpyrazole-4-carboxylic acid(24.0 g, 97%).

d) To a solution of 1-(4-chlorophenyl)-5-isopropylpyrazole-4-carboxylicacid (10.0 g, 37.78 mmol) in CH₂Cl₂ (150 mL) was added oxalyl chloride(9.90 mL, 113.6 mmol) and DMF (0.15 mL). The mixture was stirred for 2 hat room temperature, concentrated in vacuo, and re-dissolved into 100 mLof acetone. The obtained acyl chloride solution was added portionwise toanother flask containing NaN₃ (12.31 g, 190 mmol) in H₂O (100 mL) at 0°C. After 10 min, The reaction mixture was diluted with brine (300 mL)and extracted with EtOAc (500 mL). The organic layer was separated,dried over anhydrous sodium sulfate, and concentrated in vacuo. Theobtained acyl azide was diluted in 150 mL of toluene and heated at 95°C. until no more gas evolution occurred (approx. 1.5 h). The reactionmixture was cooled to rt and treated with 100 mL of dioxane and 300 mLof 1 M aqueous HCl. The resulting two-phase solution was heated to 95°C. After approx. 2.5 h, the reaction mixture was cooled to rt, madebasic with dilute NH₄OH, and extracted with EtOAc (500 ml). The organiclayer was separated, dried over anhydrous sodium sulfate, concentratedin vacuo, and purified by flash chromatography (SiO₂, 0-100%,EtOAc/CH₂Cl₂ gradient elution) to yield1-(4-chlorophenyl)-5-isopropylpyrazol-4-amine (5.7 g, 64%).

e) To a solution of2-[2-methyl-4-(trifluoromethyl)imidazole-1-yl]propanoic acid (0.035 g,0.16 mmol) in CH₂Cl₂ (2 mL) was added oxalyl chloride (0.050 mL, 0.58mmol) and DMF (1 drop). The mixture was stirred for 30 min at roomtemperature and concentrated in vacuo. The residue obtained wastransferred to another flask containing1-(4-chlorophenyl)-5-isopropylpyrazol-4-amine (0.035 g, 0.15 mmol) andNEt₃ (0.060 mL, 0.43 mmol) in CH₂Cl₂ (3 mL). After stirring for 30 minat room temperature, the reaction was quenched with saturated aqueousNaHCO₃ (30 mL) and the mixture was extracted with ethyl acetate (50 mL).The organic layer was separated, dried over anhydrous sodium sulfate,concentrated in vacuo, and purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to afford the TFA salt of thetitled compound (0.050 g, 57%) as a white solid. ¹H NMR (TFA salt) (400MHz, CD3OD) δ 7.88 (m, 1 H), 7.57 (m, 3 H), 7.40 (m, 1 H), 5.22 (q,J=7.2 Hz, 1 H), 3.00 (septet, J=7.1 Hz, 2 H), 2.51 (s, 3 H), 1.83 (d,J=7.6 Hz, 3 H), 1.19 (m, 6 H); MS: (ES) m/z calculated for C₂₀H₂₁ClF₃N₅O[M+H]⁺ 440.1, found 440.1.

Example 25 Synthesis ofN-[2-(4-Chlorophenyl)-3-isopropyl-imidazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

a) To a solution of 4-chlorobenzoyl chloride (1.75 g, 10.0 mmol) inCH₂Cl₂ (20 mL) was added Et₃N (3 mL, 21 mmol) and propan-2-amine (1.3mL, 15 mmol) at 0° C. The resulting mixture was stirred overnight atroom temperature before it was diluted with saturated aqueous NaHCO₃solution (30 mL) and extracted with EtOAc (50 mL). The organic layer wasseparated, washed with brine, dried (Na₂SO₄), filtered, and concentratedin vacuo. The crude material was used directly in the next step.

b) A mixture of the crude product from step a and thionyl chloride (20mL) was refluxed overnight. After cooling to room temperature, thereaction mixture was concentrated in vacuo. The crude material was useddirectly in the next step.

c) To a stirred solution of (1Z)-4-chloro-N-isopropyl-benzimidoylchloride (1.00 g, 4.6 mmol) in toluene (2 mL) was added2-aminoacetonitrile (0.26 g, 4.6 mmol), at 0° C. The resulting solutionwas stirred at room temperature overnight before before it was dilutedwith saturated aqueous NaHCO₃ solution (10 mL) and extracted with EtOAc(20 mL). The organic layer was subsequently washed with brine, dried(Na₂SO₄), filtered, and concentrated in vacuo. Purification by flashchromatography (SiO₂, 100% EtOAc −20% MeOH/EtOAc) gave the desiredproduct (0.50 g, 2.1 mmol, 45% yield).

d) A mixture of 2-(4-chlorophenyl)-3-isopropyl-imidazol-4-amine (0.10 g,0.43 mmol), 2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanoic acid(0.09 g, 0.43 mmol), HATU (0.16 g, 0.43 mmol), and Et₃N (200 μL, 1.4mmol) in CH₂Cl₂ (1 mL) was stirred at room temperature for 30 min. Thereaction mixture was then diluted with 10 mL of saturated aqueous NaHCO₃solution and extracted with EtOAc. The organic layer was subsequentlywashed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo.The resulting crude product was purified by flash chromatography (SiO₂,100% EtOAc −30% MeOH/EtOAc) to afford the titled compound as a whitesolid (0.046 g, 0.10 mmol, 23% yield). ¹H NMR (400 MHz, CDCl₃) 7.52 (s,1 H), 7.44 (d, J=8.0 Hz, 2 H), 7.32 (d, J=8.0 Hz, 2 H), 7.07 (s, 1 H),5.18 (q, J=7.1 Hz, 1 H), 4.36 (q, J=7.0 Hz, 1 H), 2.42 (s, 3 H), 1.78(d, J=7.1 Hz, 3 H), 1.33 (d, J=7.0 Hz, 6 H). MS: (ES) m/z calculated forC₂₀H₂₁ClF₃N₅O [M+H]⁺ 440.1, found 440.4.

Example 26 Synthesis of(2S)—N-[1-(4-chlorophenyl)-5-cyclobutyl-pyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamideand(2R)—N-[1-(4-chlorophenyl)-5-cyclobutyl-pyrazol-4-yl]-2-[2-methyl-4-(trifluoromethyl)imidazol-1-yl]propanamide

a) Pyridine (20.46 mL, 253 mmol) was added to a solution ofcyclobutanecarboxylic acid chloride (10.0 g, 84.3 mmol) andisopropylidene malonate (12.16 g, 84.3 mmol) in CH₂Cl₂ (100 mL) at 0° C.and the mixture was stirred at room temperature for 1.5 h. Methanol (100mL) was then added and the resulting mixture was stirred at reflux for 3h, cooled to room temperature and partitioned between aqueous HCl (1 M,200 mL) and EtOAc (500 mL). The organic layer was separated, dried overanhydrous sodium sulfate, concentrated in vacuo and purified by flashchromatography (SiO₂, 0-20% EtOAc/hexanes gradient elution) to givemethyl 3-cyclobutyl-3-oxo-propanoate (11.6 g, 88% yield).

b) A mixture of methyl 3-cyclobutyl-3-oxo-propanoate (5.8 g, 37.2 mmol)and N,N-dimethylformamide dimethyl acetal (25 g, 210 mmol) was stirredat 100° C. for 1 h. After cooling to room temperature the mixture wasconcentrated in vacuo to give an oily residue that was directly carriedto the next step.

c) A mixture of the intermediate (˜37.2 mmol) obtained in step b,4-chlorophenylhydrazine hydrochloride (6.67 g, 37.2 mmol), and K₂CO₃(10.3 g, 74.4 mmol) in DMF (50 mL) was stirred at 100° C. for 1 h. Aftercooling to room temperature the mixture was diluted with aqueous HCl(200 mL) and extracted with EtOAc (500 mL). The organic layer wasseparated, dried over anhydrous sodium sulfate, concentrated in vacuo,and purified by flash chromatography (SiO₂, 0-10% EtOAc/CH₂Cl₂ gradientelution) to give methyl1-(4-chlorophenyl)-5-cyclobutyl-pyrazole-4-carboxylate (8.3 g, 76%yield).

d) A mixture of methyl1-(4-chlorophenyl)-5-cyclobutyl-pyrazole-4-carboxylate (8.3 g, 28.5mmol) and lithium hydroxide monohydrate (3.6 g, 85.6 mmol) in MeOH (25mL), THF (25 mL), and H₂O (12 mL) was stirred at 80° C. for 1 h. Aftercooling to room temperature the mixture was acidified with 1 M aqueousHCl and extracted with EtOAc (400 mL). The organic layer was separated,dried over anhydrous sodium sulfate, and concentrated in vacuo to yield1-(4-chlorophenyl)-5-cyclobutyl-pyrazole-4-carboxylic acid (6.92 g, 87%yield).

e) To a mixture of 1-(4-chlorophenyl)-5-cyclobutyl-pyrazole-4-carboxylicacid (4.0 g, 14.4 mmol) in CH₂Cl₂ (100 mL) was added oxalyl chloride(3.78 mL, 43.4 mmol) and DMF (0.06 mL). After 2 h at room temperature,the reaction mixture was concentrated in vacuo, re-dissolved in 40 mL ofacetone, and added to a 0° C. solution of NaN₃ (3.75 g, 57.8 mmol) inH₂O (40 mL). Brine (150 mL) and EtOAc (350 mL) were then added. Theorganic layer was separated, dried over anhydrous sodium sulfate andconcentrated in vacuo. The residue was stirred in 100 mL of toluene at95° C. for 1 h, cooled to room temperature, and then treated with 150 mLof 6 M aqueous HCl at 110° C. for 1 h. After cooling to room temperaturethe mixture was basified with dilute NH₄OH and extracted with EtOAc (500mL). The organic layer was separated, dried over anhydrous sodiumsulfate, concentrated in vacuo, and purified by flash chromatography(SiO₂, 0-100% EtOAc/CH₂Cl₂ gradient elution) to yield1-(4-chlorophenyl)-5-cylocbutyl-pyrazol-4-amine (2.9 g, 81% yield).

f) A mixture of(2S)-2-[2-methyl-4-(trifluoromethyl)imidazole-1-yl]propanoic acid (0.046g, 0.21 mmol), 1-(4-chlorophenyl)-5-cylocbutyl-pyrazol-4-amine (0.046 g,0.18 mmol) and pyridine (0.072 mL, 0.92 mmol) in CH₃CN (1 mL) and EtOAc(1 mL) at 0° C. was treated with 1-propylphosphonic acid cyclicanhydride (50% in EtOAc, 0.24 mL, 0.4 mmol) for 15 min at 0° C., thenquenched with 0.5 M aq. HCl (10 mL), neutralized with saturated aqueousNaHCO₃ (30 mL), and extracted with ethyl acetate (100 mL). The organiclayer was collected, dried over anhydrous sodium sulfate, concentratedin vacuo, and purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to afford the TFA salt of thetitled compound. It was then converted to the free form by treating withsaturated aqueous NaHCO₃ followed by EtOAc extraction to afford thetitled compound (0.055 g, 65% yield). ¹H NMR (400 MHz, CDCl₃) δ 7.93 (s,1 H), 7.42 (m, 3 H), 7.27 (m, 3 H), 4.92 (q, J=7.3 Hz, 1 H), 3.56 (m, 1H), 2.49 (s, 3 H), 1.62-1.96 (m, 9 H); MS: (ES) m/z calculated forC₂₁H₂₁ClF₃NO₅ [M+H]⁺ 452.1, found 452.1. Chiral HPLC (Regis Pack CLA-1,cat #793104, 25 cm×4.6 mm, 5 micron; eluent: 0.1% diethylamine/IPA, 0.7ml/min) analysis of the product showed an enantiomeric ratio of 48:1.The (S)-enantiomer (major) had a retention time of 6.8 min, and the(R)-enantiomer (minor) had a retention time of 5.2 min.

Example 27

This example illustrates the evaluation of the biological activityassociated with compounds of interest of the invention.

Materials and Methods

A. Cells

1. CCR1 Expressing Cells

a) THP-1 Cells

THP-1 cells were obtained from ATCC (TIB-202) and cultured as asuspension in RPMI-1640 medium supplemented with 2 mM L-glutamine, 1.5g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, 1 mM sodiumpyruvate, 0.05% 2-mercaptoethanol and 10% FBS. Cells were grown under 5%CO₂/95% air, 100% humidity at 37° C. and subcultured twice weekly at 1:5(cells were cultured at a density range of 2×10⁵ to 2×10⁶ cells/mL) andharvested at 1×10⁶ cells/mL. THP-1 cells express CCR1 and can be used inCCR1 binding and functional assays.

2. Chemotaxis Assays

Chemotaxis assays were performed using 5 μm pore polycarbonate,polyvinylpyrrolidone-coated filters in 96-well chemotaxis chambers(Neuroprobe; Gaithersburg, Md.) using chemotaxis buffer (Hank's balancedsalt solution (HBSS) and 1% FBS). CCR1 chemokine ligands (i.e., MIP-1α,CCL15/Leukotactin; R&D Systems; Minneapolis, Minn.) are use to evaluatecompound mediated inhibition of CCR1 mediated migration. Otherchemokines (i.e., SDF-1α; R&D Systems; Minneapolis, Minn.) are used asspecificity controls. The lower chamber was loaded with 29 μl ofchemokine (i.e., 0.1 nM CCL15/Leukotactin) and varying amounts ofcompound, the top chamber contained 100,000 THP-1 or monocyte cells in20 μl. The chambers were incubated 1-2 hours at 37° C., and the numberof cells in the lower chamber quantified either by direct cell counts infive high powered fields per well or by the CyQuant assay (MolecularProbes), a fluorescent dye method that measures nucleic acid content andmicroscopic observation.

B. Identification of Inhibitors of CCR1

One of the primary functions of chemokines is their ability to mediatethe migration of chemokine receptor-expressing cells, such as whiteblood cells. To confirm that a compound of interest inhibited not onlyCCR1 specific binding and signaling (at least as determined by calciummobilization assays), but also CCR1 mediated migration, a chemotaxisassay was employed. THP-1 myelomonocytic leukemia cells, which resemblemonocytes, as wells as freshly isolated monocytes, were used as targetsfor chemoattraction by CCR1 chemokine ligands (i.e., MIP-1α,CCL15/leukotactin). Cells were placed in the top compartment of amicrowell migration chamber, while MIP-1α (or other potent CCR1chemokine ligand) and increasing concentrations of a compound ofinterest was loaded in the lower chamber. In the absence of inhibitor,cells will migrate to the lower chamber in response to the chemokineagonist; if a compound inhibited CCR1 function, then the majority ofcells will remain in the upper chamber. To ascertain a compound ofinterest's affinity for CCR1 as well as to confirm its ability toinhibit CCR1 mediated cell migration, inhibitory activity was titeredover a 1×10⁻¹⁰ to 1×10⁻⁴ M range of compound concentrations in thischemotaxis assay. In this assay, the amount of compound was varied;while cell number and chemokine agonist concentrations were heldconstant. After the chemotaxis chambers were incubated 1-2 hours at 37°C., the responding cells in the lower chamber were quantified bylabeling with the CyQuant assay (Molecular Probes), a fluorescent dyemethod that measures nucleic acid content, and by measuring with aSpectrafluor Plus (Tecan). The computer program Prism from GraphPad,Inc. (San Diego, Ca) was used to calculate IC₅₀ values. IC₅₀ values arethose compound concentrations required to inhibit the number of cellsresponding to a CCR1 agonist by 50%.

1. In Vivo Efficacy

a) Rabbit Model of Destructive Joint Inflammation

A rabbit LPS study was conducted essentially as described in Podolin, etal. J. Immunol. 169(11):6435-6444 (2002). Female New Zealand rabbits(approximately 2 kilograms) were treated intra-articularly in both kneeswith LPS (10 ng). The compound of interest, for example 1.016,(formulated in 1% methocel) or vehicle (1% methocel) was dosed orally ata 5 ml/kg dose volume at two times (2 hours before the intra-articularLPS injection and 4 hours after the intra-articular LPS injection).Sixteen hours after the LPS injection, knees were lavaged and cellscounts were performed. Beneficial effects of treatment were determinedby reduction in the number of inflammatory cells recruited to theinflamed synovial fluid of the knee joints. Treatment with the compoundof interest resulted in a significant reduction in recruitedinflammatory cells.

b) Evaluation of a Compound of Interest in a Rat Model of CollagenInduced Arthritis

A 17 day developing type II collagen arthritis study is conducted toevaluate the effects of a compound of interest on arthritis inducedclinical ankle swelling. Rat collagen arthritis is an experimental modelof polyarthritis that has been widely used for preclinical testing ofnumerous anti-arthritic agents (see Trentham, et al., J. Exp. Med.146(3):857-868 (1977), Bendele, et al., Toxicologic Pathol. 27:134-142(1999), Bendele, et al., Arthritis Rheum. 42:498-506 (1999)). Thehallmarks of this model are reliable onset and progression of robust,easily measurable polyarticular inflammation, marked cartilagedestruction in association with pannus formation and mild to moderatebone resorption and periosteal bone proliferation.

Female Lewis rats (approximately 0.2 kilograms) are anesthetized withisoflurane and injected with Freund's Incomplete Adjuvant containing 2mg/mL bovine type II collagen at the base of the tail and two sites onthe back on days 0 and 6 of this 17 day study. A compound of interest isdosed daily in a sub-cutaneous manner from day 0 till day 17 at aefficacious dose. Caliper measurements of the ankle joint diameter aretaken, and reduced joint swelling is taken as a measure of efficacy.

Murine Model of Dermatological Disease

Compounds of the invention can be assessed in the murine model of dermaldelayed type hypersensitivity induced by oxazolone. Briefly, 8-10 weekold BALB/c mice are sensitized topically with a 1% solution of oxazolonedissolved in ethanol on their shaved abdomens on day 0. On day 6 postsensitization mice are dosed orally with either vehicle or increasingdoses of a compound of the invention immediately prior to and 4 hoursfollowing a topical challenge with a 0.5% solution of oxazolone inethanol on the right ear. The following day (day 7), ear thicknesses aremeasured using caliper measurements. Animals treated with compound havesignificantly reduced ear swelling compared to vehicle treated controlsindicating a compound mediated decrease in oxazolone induced dermalhypersensitivity.

Murine Asthma Model

Compounds of the invention can be assessed in the murine model ofallergic asthma. Asthma is induced in 8-10 week old BALB/c mice bysensitizing mice with OVA in Alum adjuvant on days 0 and 10. On day 20mice are challenged with OVA in PBS intranasally to elicit airwayinflammation. Groups of mice are either treated with vehicle, orincreasing doses of a compound of the invention starting on day 20 andlasting until day 23. Animals are analyzed at day 23 after theintranasal OVA challenge for cellular infiltrates in bronchoalveolarlavage (BAL). A significant reduction in BAL leukocyte numbers relativeto vehicle treated mice indicates the compound is effective in thismodel.

Murine Model of Systemic Lupus Erythematosus

This example describes a procedure to evaluate efficacy of CCR1antagonists for treatment of Systemic Lupus Erythematosus (SLE). FemaleNZB/W FI mice spontaneously develop an SLE-like pathology commencing at6 months of age that is characterized by proteinuria, serumautoantibodies, glomerulonephritis, and eventually death. Three seriesof NZB/W FI mouse groups comprising 20 mice per group are tested forefficacy of CCR1 antagonist as follows: One series of mice additionallyreceives phosphate buffered saline (PBS) and Tween 0.5% i.p. soon afterweaning, and thereafter at varying dosing schedules. A second seriesconsists of groups of mice receiving different doses of the CCR1antagonist given either intra-peritoneally, intra-venously,sub-cutaneously, intramuscularly, orally, or via any other mode ofadministration soon after weaning, and thereafter at varying dosingschedules. A third series of mice, serving as positive control, consistsof groups treated with anti-IL10 antibodies given soon after weaning,and thereafter at varying dosing schedules. Disease development ismonitored in terms of eventual mortality, kidney histology, serumautoantibody levels, and proteinuria.

Murine Model of Cancer

This example describes a procedure to evaluate efficacy of CCR1antagonists for treatment of malignancy. Normal mouse strains can betransplanted with a variety of well-characterized mouse tumor lines,including a mouse thymoma EL4 which has been transfected with OVA toallow easy evaluation of tumor specific antigen responses followingvaccination with OVA. Three series of mouse groups from any of thesetumor models are tested for CCR1 antagonist efficacy as follows: Oneseries of mice additionally receives PBS and Tween 0.5% i.p. soon aftertumor transplant, and thereafter at varying dosing schedules. A secondseries consists of groups of mice receiving different doses of the CCR1antagonist given either intra-peritoneally, intra-venously,sub-cutaneously, intramuscularly, orally, or via any other mode ofadministration soon after tumor transplant, and thereafter at varyingdosing schedules. A third series of mice, serving as positive control,consists of groups treated with either anti-IL4 antibodies, anti-IFNgantibodies, IL4, or TNF, given i.p. soon after tumor transplant, andthereafter at varying dosing schedules. Efficacy is monitored via tumorgrowth versus regression. In the case of the OVA-transfected EL4 thymomamodel, cytolytic OVA-specific responses can be measured by stimulatingdraining lymph node cells with OVA in vitro, and measuringantigen-specific cytotoxicity at 72 hours.

Murine Model of Psoriasis

This example describes procedures to evaluate the efficacy of CCR1antagonists in psoriasis. A rodent model of psoriasis can be obtained byintra-venously transferring a population of purified T cells (designatedCD45Rbhi T cells) obtained from the spleens of BALB/c mice intoimmunodeficient recipient CB.17 scid/scid mice. Mice develop signs ofredness, swelling, and skin lesions resembling those of human psoriasisin their ear, feet and tail by 8 weeks after transfer. Three series ofmouse groups, comprising 10-15 CB.17 scid/scid mice per group, areinjected with purified CD45Rbhi T cells. One series of mice additionallyreceives phosphate buffered saline (PBS) and Tween 0.5% i.p. at theinitial cell transfer, and at different dosing schedules thereafter. Asecond series consists of groups of mice receiving different doses ofthe CCR1 antagonist given either intra-peritoneally, intra-venously,sub-cutaneously, intra-muscularly, orally, or via any other mode ofadministration at the initial cell transfer, and at different dosingschedules thereafter. A third series of mice, serving as positivecontrol, consists of groups treated with antibodies to either IL-12,IL-4, IFNg, or TNF, or with cytokine IL-10 at the initial cell transfer,and at different dosing schedules thereafter. Animals are monitored fordevelopment of psoriatic-like lesions for 3 months after cell transfer.

Murine Model of Inflammatory Bowel Diseases

The MDR1a-knockout mice, which lack the P-glycoprotein gene,spontaneously develop colitis under specific pathogen-free condition.The pathology in these animals has been characterized as Th1-type Tcell-mediated inflammation similar to ulcerative colitis in humans.Disease normally begins to develop at around 8-10 weeks after birth.However the ages at which disease emerges and the ultimate penetrancelevel often vary considerably among different animal facilities. In astudy using the MDR1a-knockout mice, a CCR1 antagonist can be evaluatedprophylacticly or therapeutically depending on time of administration.Female mice (n=34) are dosed with a compound of interest as appropriateto the compound eg daily in a sub-cutaneous manner at a efficaciousdose. The study is evaluated for IBD associated growth retardation andscoring of anal discharge and irritation. A compound which reduces analdischarge and irritation or inhibits IBD associated growth retardationindicates efficacy of compound in this indication.

Murine Model of Solid Tumors

The mouse RENCA tumor model accurately mimics the progression of humanadult renal cell carcinoma specifically with reference to spontaneousmetastasis to lungs and serves as a model for solid tumors. Balb/c 6-8week old female mice are inoculated with approximately 5e5 RENCA cells(mouse renal adenocarcinoma; ATCC cat# CRL-2947) under the kidneycapsule and kidney tumor growth is observed over 22 days, with lungmetastasis observed as early as day 15. Animals are dosed with eithervehicle or a compound of the invention eg daily subcutaneously, from thetime of tumor implantation to monitor effects on primary growth, or at alater time (eg day 7) to monitor the compound effect on metastasis.Primary tumor areas are measured twice a week using mechanical calipers.Tumor volumes are calculated by the formula v=pab2/6, where a is thelongest diameter and b is the next longest diameter perpendicular to a.A reduction in tumor volume or incidence of metastasis indicatesefficacy of compound in this indication.

Murine Model of Inflammation

A method of inducing peritoneal inflammation by the introduction of 3%thioglycolate into the peritoneum is well know in the art. Following theintroduction of thioglycolate, a rapid influx of immune cells to thesite, primarily CCR1 bearing neutrophils, results in local inflammationat 24 hours. A peritoneal exudate can be sampled, and the cell numberand composition can be assessed to determine the anti-inflammatoryproperties of a compound of interest administered before, during orafter the thioglycolate induction.

In Table 1 (below), structures and activity are provided forrepresentative compounds described herein. Activity is provided asfollows for the chemotaxis assay as described above: +, 10 μM>IC₅₀>100nM; ++, IC₅₀≤100 nM.

TABLE 1 Migration IC50 (nM) 1.001

++ 1.002

+ 1.003

+ 1.004

+ 1.005

+ 1.006

+ 1.007

+ 1.008

+ 1.009

++ 1.010

++ 1.011

++ 1.012

++ 1.013

+ 1.014

+ 1.015

++ 1.016

++ 1.017

++ 1.018

++ 1.019

++ 1.020

+ 1.021

+ 1.022

++ 1.023

+ 1.024

+ 1.025

+ 1.026

+ 1.027

+ 1.028

+ 1.029

++

What is claimed is:
 1. A pharmaceutical composition comprising apharmaceutically acceptable excipient or carrier and a compound ofFormula (Ib2)

wherein each A is independently selected from the group consisting of Nand CH; A¹ is N or C(R⁵); A² is N or C(R⁷); R¹ is selected from thegroup consisting of H, halogen, CN, C₁₋₈ alkyl, C₁₋₈ haloalkyl,—CO₂R^(a)and —SO₂R^(a); R³ is a member selected from the group consisting of H,halogen, CN, C₁₋₈ alkyl, C₃₋₈ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl,C₁₋₈ haloalkyl, C₁₋₈ hydroxyalkyl, —OR^(a), —CO₂R^(a), —NR^(a)R^(b), and—CONR^(a)R^(b), aryl, 5- or 6-membered heteroaryl, and R⁵, R⁷ and R⁸ areeach independently selected from the group consisting of H, halogen, CN,C₁₋₈ alkyl, C₃₋₈ cycloalkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₁₋₈ haloalkyl,C₁₋₈ hydroxyalkyl, —OR^(a), —CO₂R^(a), —NR^(a)R^(b), and —CONR^(a)R^(b);each R^(a) and R^(b) is independently selected from the group consistingof hydrogen, hydroxyl, halogen, cyano, C₁₋₈ alkyl, C₁₋₈ alkoxy,C₁₋₈haloalkyl, C₃₋₆ cycloalkyl, C₃₋₆ cycloalkylalkyl, amino, C₁₋₈alkylamino, di C₁₋₈ alkylamino, carboxamide, carboxy C₁₋₄ alkyl ester,carboxylic acid, and —SO₂—C₁₋₈ alkyl, or a pharmaceutically acceptablesalt thereof.
 2. A pharmaceutical composition of claim 1, wherein R¹ isCl or F; R³ is selected from the group consisting of C₁₋₈ alkyl, C₁₋₈haloalkyl, C₁₋₈ hydroxyalkyl, wherein the alkyl portions of R³ areoptionally further substituted with 1-3 R^(a).
 3. A pharmaceuticalcomposition of claim 1, wherein the ring portion having N, A^(l) and A²as ring vertices is selected from the group consisting of:

wherein R⁷ is H or Cl, and R⁸ is C₁₋₈ alkyl optionally substituted with1 or 2 R^(a).
 4. A pharmaceutical composition of claim 1, wherein thecompound of Formula (Ib2) has the Formula (Ib3)


5. A pharmaceutical composition of claim 4, wherein R¹ is Cl or F.
 6. Apharmaceutical composition of claim 1, wherein said compound is selectedfrom the group consisting of:

or a pharmaceutically acceptable salt, hydrate, solvate, rotamer orN-oxide thereof.
 7. A pharmaceutical composition of claim 1, whereinsaid compound has the formula:

or a pharmaceutically acceptable salt thereof.
 8. A pharmaceuticalcomposition of claim 1, wherein said compound has the formula:

or a pharmaceutically acceptable salt thereof.